text:
Study of Vaccination With Autologous Acute Myeloblastic Leukemia Cells in Patients With Advanced Myelodysplasia or Acute Myelogenous Leukemia
- Full Text View
- Tabular View
- No Study Results Posted
- Disclaimer
- How to Read a Study Record
|Acute Myelogenous Leukemia Myelodysplasia
|Biological: autologous tumor cells
|Phase 1
|Study Type:
|Study Design:
|Allocation: Non-Randomized
Intervention Model: Parallel Assignment
Masking: Open Label
Primary Purpose: Prevention
|Official Title:
|A Phase I Study of Vaccination With Lethally Irradiated, Autologous Acute Myeloblastic Leukemia Cells Engineered by Adenoviral Mediated Gene Transfer to Secrete Human Granulocyte-Macrophage Colony Stimulating Factor in Patients With Advanced Myelodysplasia or Acute Myelogenous Leukemia
- To determine the feasibility of preparing lethally irradiated autologous myeloblastic leukemia cells engineered by adenoviral mediated gene transfer to secrete GM-CSF in patients with myelodysplastic syndromes (MDS) or AML
- To determine the safety and biologic activity of vaccination with lethally irradiated, autologous myeloblastic leukemia cells engineered by adenoviral mediated gene transfer to secrete GM-CSF in patients with MDS or AML
|Estimated Enrollment:
|Study Start Date:
|January 2000
|Study Completion Date:
|March 2006
|Primary Completion Date:
|March 2006 (Final data collection date for primary outcome measure)
This study will make use of leukemic myeloblasts harvested by bone marrow aspirate in patients with myelodysplasia and acute myelogenous leukemia. These harvested tumor cells will be modified by adenoviral mediated gene transfer to secrete granulocyte-macrophage colony stimulating factor (GM-CSF).
Patients will be administered vaccines at one of three dose levels (as determined by total cell yield). Vaccinations will be given weekly for three weeks, followed by every other week until the vaccine supply is exhausted or when patients are removed from the study.
The patient will receive a minimum of six vaccinations, but more will be administered if the vaccine is available.
During the course of the study, patients will be tested to see how their immune system is reacting to the vaccinations. Testing will include bloodwork evaluating the immune cells in the body at monthly intervals. Skin biopsies may also be performed to see if an immune reaction is occuring at the injection site.
During the first course of treatment, a bone marrow biopsy and aspirate may be performed monthly.
The length of time on this study depends upon the number of vaccines available and whether or not unacceptable side effects occur.
|Ages Eligible for Study:
|18 Years and older (Adult, Senior)
|Sexes Eligible for Study:
|Accepts Healthy Volunteers:
Inclusion Criteria:
- Patients must have pathologically documented myelodysplasia or acute myelogenous leukemia.
- The patients with myelodysplasia must also have: French-American-British (FAB) subtype refractory anemia with excess blasts (RAEB) or refractory anemia with excess blasts in transformation (RAEB-T), or normal or hypercellular bone marrow.
- The patients with acute myelogenous leukemia must also: not be candidates for myelosuppressive chemotherapy due to age or comorbid disease, or have relapsed acute myelogenous leukemia or be refractory to standard therapy and not likely to require cytoreductive therapy within 60 days
- Eastern Cooperative Oncology Group (ECOG) performance status 0 or 1.
- Estimated life expectancy of 6 months or greater.
- Age at least 18 years.
- Greater than 4 weeks from any chemotherapy, radiotherapy, immunotherapy, or systemic glucocorticoid therapy (non-glucocorticoid hormonal therapy allowed).
- Greater than 2 months following bone marrow or peripheral blood stem cell transplantation or treatment with donor lymphocyte infusion (DLI).
Exclusion Criteria:
- Uncontrolled active infection.
- Pregnancy or nursing mothers.
- Previous participation in an adenovirus based trial.
- The patients with myelodysplasia who have either: FAB subtype refractory anemia (RA), refractory anemia with ringed sideroblasts (RARS), chronic myelomonocytic leukemia (CMML), or the presence of hypocellular bone marrow.
- Chemotherapy, radiotherapy, immunotherapy, or systemic steroid therapy within the last 4 weeks.
- Active central nervous system (CNS) disease.
- Evidence of infection with the human immunodeficiency virus.
- Active psychiatric or mental illness making informed consent or careful clinical follow-up unlikely.
Please refer to this study by its ClinicalTrials.gov identifier: NCT00136422
|United States, Massachusetts
|Dana-Farber Cancer Institute
|Boston, Massachusetts, United States, 02115
|Principal Investigator:
|Daniel J. DeAngelo, MD, PhD
|Dana-Farber Cancer Institute
|ClinicalTrials.gov Identifier:
|NCT00136422 History of Changes
|Other Study ID Numbers:
|Study First Received:
|August 25, 2005
|Last Updated:
|March 9, 2011
Keywords provided by Dana-Farber Cancer Institute:
acute myelogenous leukemia
Additional relevant MeSH terms:
Leukemia, Myeloid
Leukemia, Myeloid, Acute
Myelodysplastic Syndromes
Neoplasms by Histologic Type
Bone Marrow Diseases
Hematologic Diseases
Precancerous Conditions
ClinicalTrials.gov processed this record on June 23, 2017


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Rustic Mirror
Rebuilding Exchange
Saturday, December 15, 2012 from 11:00 AM to 2:00 PM (CST)
Who's Going
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3 of your friends are going. Share it to get more people on board! Share
Sara Haughty
San Francisco, California
San Francisco, California
Luke Picard
London, United Kingdom
London, United Kingdom
Rick Vespucio
Florence, Italy
Florence, Italy
Event Details
Students will pick a salvaged mirror and then build their own frame from reclaimed pine to fit the mirror.
Have questions about Rustic Mirror? Contact Rebuilding Exchange
When & Where
Rebuilding Exchange
1740 W Webster
Saturday, December 15, 2012 from 11:00 AM to 2:00 PM (CST)
Add to my calendar
Rebuilding Exchange
Our mission is to create a market for reclaimed building materials. We do this by diverting materials from landfills and making them accessible for reuse through our retail warehouse, by promoting sustainable deconstruction practices, by providing education and job training programs, and by creating innovative models for sustainable reuse.
Please note our new location! 1740 W Webster Ave., Chicago IL
Rustic Mirror
Things to do in Chicago, IL

subset: commoncrawl


text:
Roy Price Makes Sure Your Script Gets Read at Amazon Studios
Price oversees the development and production of original shows and films for Amazon Studios, the production platform launched in 2010 by the Seattle-based e-commerce giant.
Roy Price Makes Sure Your Script Gets Read at Amazon Studios
Price oversees the development and production of original shows and films for Amazon Studios, the production platform launched in 2010 by the Seattle-based e-commerce giant.
'SingleDumb' Spotlights the Not-So-Good, the Bad, and the Ugly of Online Dating
Though many keep dating horror stories to themselves, Alexandra Kern is sharing hers with the world via her comedic Web series, "SingleDumb."
Web Writer-Producer Freddie Wong On Using Technology To Get Ahead
USC grad Freddie Wong built a strong Internet presence and following for his Web series "Video Game High School" and, with Matt Arnold, launched the production company Rocket Jump, where he continues to innovate content delivery.
9 Tips for Marketing Your Web Series
Creating successful Web content isn't just about the writing and filming. If your marketing is all wrong, your series can totally flop!
'Llama Cop': The Llama-Starring Web Series You Never Knew You Needed
"Llama Cop" is the quintessential buddy cop Web series, except for one small detail.
Understudies Step Into the Spotlight in a New Web Series
Created and written by real-life former understudy Elisabeth Gray, Web series "Understudies" follows one plucky actor as she covers the lead role in the vampire adaptation of "Breakfast at Tiffany's," "Twilight at Tiffany's."
'Preggers' Web Series Creators on Being Proactive Between Gigs
Eilis Cahill and Catherine Decioccio tell us about their award-winning Web series "Preggers" and how it kept them and their friends busy between jobs.
'Mythos' Creator Miriam Pultro on How to Shoot a Web Series on a Shoestring
Pultro speaks about the award-winning Web series following gods in the modern world—and the most important thing she did to make sure it got made!
10 Questions With…Saffron Burrows
The "Mozart in the Jungle" actor talks to Backstage about the one topic you probably shouldn't bring up in the audition room, the lengths to which she once went to prepare for a role (it involves trespassing), and her plan B if acting doesn't work out.
Starz Digital Picks Up Web Series 'Lizzie Bennet Diaries' and 'Emma Approved'
Pemberley Digital's Emmy-winning adaptation of "Pride and Prejudice," "The Lizzie Bennet Diaries," is set to be released on iTunes after being acquired by Starz Digital Media, while Season 3 of "Emma Approved" is now airing.


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Commentary: Salt Lake City did it the right way
SALT LAKE CITY — We've seen the sad pictures: weed-covered Olympic venues that were quickly of no use after their 2½ weeks of glory, largely abandoned after costing billions to build.
Then there's Salt Lake City, where the legacy of the 2002 Winter Games carries on vibrantly more than a decade later. It's a lesson all cities bidding for big-time sporting events should heed.
"We view legacy not so much as memorials and museums," said Colin Hilton, president and CEO of the Utah Olympic Legacy Foundation. "For us, it's a living legacy."
The Olympic oval in suburban Kearns, where speedskating was held during the Salt Lake City Games, is still a valued part of the community. So is the Utah Olympic Park, home of the bobsled and luge runs as well as the towering ski-jumping hills. Both are hosting U.S. Olympic trials this weekend, continuing to provide a dynamic winter sports legacy.
In fact, every venue used during the last Olympics in the United States remains fully functional, open to everyone from world-class athletes to locals who can barely stand on a pair of skates.
What a contrast with other cities.
Take Athens, which hosted the Summer Games two years after Salt Lake City. Many of those facilities were largely abandoned after the Olympics, quickly of no use to a country that found itself on the verge of financial collapse. The images of crumbling stadiums should've provided a stern warning to other cities, but no one seemed to notice. Beijing spent billions for its coming-out party in 2008, leaving behind an impressive national stadium, the Bird's Nest, that goes largely unused except as a tourist attraction.
Sochi is preparing to host the Winter Olympics in February after doling out more than $50 billion — far more than any other Olympics, winter or summer — to essentially build every venue from scratch.
Rest assured, there's no chance of Sochi getting more than kopek on the ruble for its investment. In fact, it seems very likely that five ice arenas and the main stadium — all clustered together along the Black Sea, far from the city center — will become a complex of white elephants as soon as the torch is extinguished.
Not so in Salt Lake City, which skipped the quest for gaudy architectural achievements to assure its venues would remain an important part of the community for years to come.
"We're four times busier now than we were ahead of the games," Hilton said. "There's not too many Olympic cities that can talk about being busier after the games."
It's not surprising that many of the athletes competing in various sports for spots on the U.S. Olympic team were growing up in this area during the 2002 Salt Lake City Games. Salt Lake City is already exploring a bid for another Winter Games. Hilton estimates it would cost around $2.5 billion to bring the Olympics back to Utah, a far cry from the boondoggle in Sochi.
"We love our winter sports," Hilton said. "We're active with our winter venues. Some day, if the Winter Games look to come back to us, we've got the infrastructure and citizens to host another great games."


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Check out these 2 videos on the Load Development Process: ow.ly/4naydD #SniperForum https://t.co/RXRV7ZU9iW
Bigfoot Gun Belts is a Supporting Vendor! ow.ly/4mWzSX #FNforum https://t.co/AdVK0mrx8k
Practicing with night vision optics: ow.ly/10q2aV #SniperForum https://t.co/82VdrdIIaW
Some thoughts on #USMartialLaw and rifle confiscation: ow.ly/10mY7z #prepperforums
The Forum Foundry Outdoor Forum Network includes some of the premier online communities in the Hunting, Fishing, Archery and Firearms world! These online communities are geared towards the person with the outdoor enthusiast lifestyle.


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German Shepherd Dog Forums (http://www.germanshepherds.com/forum/)
- General Puppy Stuff (http://www.germanshepherds.com/forum/general-puppy-stuff/)
- - Outdoor dog toys or entertainment? (http://www.germanshepherds.com/forum/general-puppy-stuff/432817-outdoor-dog-toys-entertainment.html)
Outdoor dog toys or entertainment?
I am finally getting my backyard fenced.. doing it myself after work so it's going to be another few weeks.
I was just curious what kind of toys or apparatus for outside entertainment is available to buy or make.. I will make a flirt pole but was also wondering if there was something that would keep her busy enough that I could maybe leave her at home for a few hours at a time. At this point she goes everywhere with me and waits in the car while I am working.
We have a neighbor that built a larger fenced in area for her dog, tucked a bit away from the house. She did it to allow more exercise for her beagle. I told her when she was planning it that if you aren't there with the dog all it will do is sit by the gate and wait to be taken back toward the house, or at least on a walk with the owner. That's all the dog does, she does not use the enclosure for anything now.
Just saying, a fenced in yard may well contain the dog, but they want to be with the family, for the most part. Unless someone is there with the dog I wouldn't expect much exercise.
I loved having a couple of sections of my yard fenced. I can let one or two girls in Far Field -- a little fenced field on the one side of my house -- while I clean kennels. And when there is grass in Near Field, a few can run and play there as well.
Also, I can throw a toy for them in the area, and I don't have to worry about anyone getting carried away and going down into the ravine and river, or out in the road.
I suppose you can purchase or build agility equipment to work with the dog on. But I would not leave the dog in there with the equipment unsupervised -- mine like the A-Frame a little too much, and if they do get hurt, I want to know how they managed it if at all possible.
half deflated soccer ball! if you're there to kick it around for her
Two of my dogs are in the back yard while I'm at work. I normally only leave hard Kong toys in the back yard for them to play with. I don't want a bored dog chewing and swallowing toys.
In the summer, I'll put out the horse trough for them to play in. I tried the kiddie pool, but they didn't care for it. I'm not sure why. I brought up a large horse trough from the barn and they both jump in and out of it...or just sit in it.
I did hang a toy once on a bungee type cord from a tree limb. At the end of the cord, I put a Kong toy. Watching them play with it I got far too worried that someone was going to either hang themselves, or end up chewing through the rope and eating the rope. Just didn't work for me. I do hang out the toy when I'm home, and they'll play with it.
If you had a camera and put it in your back yard, you'd find out that most of the day your dog will sleep.
I'm going to get a plastic swimming pool for my pup this summer. I live in the desert, so it will be hot every day. You could also set up a sandbox or area for your pup to dig. You will have to teach the dog how to use the area first, though (by burying toys, treats, and bones and encouraging the dog to find them.) Our dogs have always played outside by themselves. Most of our dogs spent equal time inside and outside, so they did entertain themselves outdoors. I don't leave my puppy outside when I leave the house yet, but I plan to for short periods of time.
Thank you everyone for your replies.. the biggest reason I want the yard is to play while I am home without worrying she will chase the cat onto the neighbors porch or run into the road. (I live in town, sadly) BUT.. if it's ever possible for her to stay home, just half the day, that would be fantastic. Kiddie pool is totally worth a try! Thanks everybody!
At country stores, you can get very large ball toys made for horses and dogs. You can also buy or make a small children's playground with slides, ladders, ropes, and whatever else you might want to add. My boy loves playgrounds.
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Australian beer hopes to boldly go into space
By Pauline Askin
SYDNEY (Reuters Life!) - Wherever man has ventured, beer has followed. Now, two Australian entrepreneurs hope that will include space.
Looking ahead to a future of growing space tourism, they have developed a full-bodied brew they believe can stand up to the trials of imbibing in space, including swelling tongues that diminish the sense of taste.
"It's going to be the first beer that will be specifically designed to be drunk in zero gravity with upcoming space tourism," said Jaron Mitchell, who owns the 4 Pines Brewing Company, a microbrewery.
Mitchell and Jason Held, at space engineering firm Saber Astronautics Australia, have developed the Vostok 4 Pines Stout, which they aim to take where no beer has gone before.
They believe they are ahead of the curve, noting that the Russians -- known to enjoy the occasional tipple -- are already taking rich tourists into space and British businessman Richard Branson is building the infrastructure for his own space tourism business, Virgin Galactic.
The two decided that a full-bodied, flavorsome, stout-style beer would work best to counter the loss of taste sensation that takes place when the tongue swells in space.
"Your face puffs out a little bit, your tongue swells up a little bit -- it's not extreme, but kind of like having a bad head cold," Held said.
"So we wanted to have a flavor that would be strong enough, that would punch through that." Continued...


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Gone To The Dogs At Combest Pool In Owensboro This Sunday
The Owensboro Parks and Recreation Department will be hosting the eighth annual "Gone to the Dogs" event at Combest Pool on Sunday, August 5th. The open swim lessons for dogs and their owners will be from 6pm-8pm. In case of inclement weather, the event will take place on Monday, August 6th. Dogs of all sizes are welcome; admission to the pool is $7 per dog and his or her owner gets in FREE!


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All week we'll be counting down the end of 2012 by counting down the Indy's top five blog posts of the year. It's a pretty big deal, we know, and that's why we're stringing it out over five days.
Without any further ado, here's No. 3:
In his only interview on the subject, Jamie Olson told the Independent that critics had it all wrong when it came to the trapping photos he posted online. Matthew Frank wrote about the controversy first on the Indy blog Nov. 2 and, in addition to Olson's quotes, posted a number of graphic images that showed Olson's dogs with a trapped coyote. Frank wrote:
Olson, reached by phone yesterday, acknowledged he made a "big-ass mistake"—not in his coyote hunting practices, but in letting the photos, which he says are more than five years old, be publicly accessible on his Facebook account.
"The reality of it," Olson said, "is that those were private pictures—personal pictures taken on personal time—and most of them were before I even worked for the agency I work for, and those were not intended for public access." He said he intended only for his dad to view the photos, so his dad could see the dog Olson was training.
Now, as a result of the photos going viral, Olson said "shit's hitting the fan and I'm having to explain things." He has since deactivated his Facebook and Twitter accounts.
Olson, a federal Wildlife Services employee in Wyoming, is under investigation and no longer giving interviews. Two congressmen have also called for an audit of Wildlife Services.
Coyotes weren't the only animal that caught our online readers attention this year. Two other wildlife posts ranked just outside of our year's top five. Back in March we posted "How to eat a wolf," which linked to a Boise Weekly story about the culinary possibilities of the canine. And in January, Alex Sakariassen had the story (and photos) of a bear that decided to den beneath the den of a Georgetown Lake cabin.
Check in tomorrow for No. 2 in our countdown.
Past entries:
No. 4: Tester wins and an election judge breaks the law
No. 5: Montana's love of beer climbs the national ranks


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HT4100 4.1, 4.5, 4.9 Discussion, Strange problem Seville 90 in Cadillac Engine Discussion; My name is Lars and i live in Sweden,3 months ago i bought this car. The car dident run so ...
- 12-06-09 04:59 AM #1Cadillac Owners Member
- Automobile(s): Seville 1990
- Join Date
- Oct 2009
- Age
- 54
- Posts
- 3
Strange problem Seville 90
My name is Lars and i live in Sweden,3 months ago i bought this car.
The car dident run so well but i tought it would be some minor problems,the
car has only run 80,000 miles and are in good condition.
I changed plugs,cables ,distributorcap,rotor,ignitioncoil oil and filters.
The car stills run badly so i have to go deeper and i found out that i have two
bad fuel injectors.I orderd 8 new Bosch injectors from fiveomotors becourse they are to expensive in Sweden and i changed the fuel pressure regulator as
well becourse it dident seem to work properly.The fuel pressure is 42 psi.
Now when i have explained what i have done with the car here is the problem
When i start the engine and she is cold it runs well but when the car is in normal operating temperature she begins to run badly it feels like there is not
the power their should be in the engine,but when i drive the car about 10 miles she starts run wery nice and then i can drive her 100 miles and there is no problem.The next problem is when i stop the engine for about 5 minutes she is wery hard to start,and i have to drive her for about 10 miles and she runs very good again.I hope you can anderstand me and maybe give me a little help.Thanks
Re: Strange problem Seville 90
Do you have the FSM for the car? If not:
From the description: it runs ok after extended warm-up? Just a guess, but maybe the problem is that it's over-cooling. Bad/wrong T-Stat and /or bad/wrong Radiator cap....
Any trouble codes? Knowing how to access all the info in the computer(not just the trouble codes)would be the best clues to the detective story you have on your hands. The FSM has all the tests/procedures to help solve problems. Best tool you can own.
BTW: not familiar with 5motors. Ever looked at RockAuto dot com for Cadillac parts?
Re: Strange problem Seville 90
Thanks for your answer!I forgot to say that i have a new radiator and cap,the
only code i have is B120 and i think it´s twilight sentinel.I also have a new t-stat and when the engine is warm the temperature is about 195 degr.F.
I couldent find a shop manual for the 90 Seville on Ebay.
I am thankfull for all your help,and i am sorry that i dont now the right words for everything i hope you can anderstand what i mean.
Re: Strange problem Seville 90
The link I put up has over 20 1990 Cadillac Seville Factory Service Manuals!
Click it.....
Re: Strange problem Seville 90
I am sorry to say that i dident understand that it is the same manual for
Eldorado and Seville.but thanks for your help.


text:
Today's Devotional
Far Greater LoveWhat does it look like when you love the world more than you love God?
Let's Make ItFacebook official!
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I was never taught about God but found Him anyway. I've been a prodical more times than I care to admit but I've come back for good this time and I'm not going anywhere. I'm by no means where I want to be in my walk with my Lord but I'm getting closer everyday.
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State Dept. Report Ammo for XL Pipeline Advocates
State Dept. Report Ammo for XL Pipeline Advocates
145 results for 'advocate'. Showing results 37 - 54
State Dept. Report Ammo for XL Pipeline Advocates
Senate Bill Would Curb Access to Student Data
DuBois Felt Alone Advocating for Religious Liberty
Cleveland Browns lineman Garrett Gilkey believes that no matter how old or big you are, bullying is never acceptable.
Young Duck Dynasty Fan Makes Pitch for Shoes
Traditional Marriage Backers: Court Set Bad Precedent
Critics: Oregon Bill a 'Sex Trafficking Magnet'
Pro-Lifers, Pro-Choicers Weigh in on Gosnell Trial
Obama Makes Historic Speech to Planned Parenthood
A Raw Deal? Federal Gov't Targets Oyster Farm
The government stands accused of using bogus conservationist research to close a popular California business. Now it's a life...
Americans Warned: Home Schoolers Stripped of Rights
Islamic Extremism Spreading Across Africa
British lawmakers voted, Tuesday overwhelmingly in favor of a bill to legalize same sex marriage. The legislation is championed...
The special needs of children with disabilities are often ignored in developing countries. That's why a U.S. ministry is...
Texas, Calif. Activists hold 'March for Life' Rallies
Thai Orphans Benefiting from Family-Based Care
Child advocates say it's a tragedy: thousands of Thai kids growing up without a parent, separated from their culture....


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My dad has died: how do I change the ownership of the car?
Probate answer:
To change the ownership of the car, this is the official line.
"What to do with the V5C if the registered keeper of the vehicle dies
The registered keeper's legal representative must fill in section 6 of the V5C
with their details or the details of the person who will now be responsible for
the vehicle and send this to us. If there is a new keeper, the new keeper must
also sign the V5C.
To prevent any unnecessary correspondence which may cause distress,
please send a covering letter with the V5C. The covering letter should give the
vehicle's registration number, the name and address of the registered keeper
who has died and the date they died. Please send the letter and V5C to:
Sensitive Casework
You do not need to send in the death certificate."
In effect, this means that there may be difficulty establishing who should in fact be the legal owner, as only the "personal representative" – normally, the person appointed as executor in the Will or the person appointed under Letters of Administration. however, dealing with this issues could mean that you have appointed yourself accidentally as administrator of the whole estate with the consequent duties a liabilities.


text:
Free personalized radio that
plays the music you love
Features of This Trackelectronica influences
repetitive melodic phrasing
major key tonality
a dynamic male vocalist
prominent percussion
emphasis on instrumental arranging
and many other similarities identified in the Music Genome Project
These are just a few of the hundreds of attributes cataloged for this track by the Music Genome Project.show more
Similar Tracks
This is the perfect song with 2001: Space Odyssey clips paired to it. Some guy put it on YouTube and now the song always gives me that vibe.
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LME49810 on ebay
1 Attachment(s)
Hi guys, I noticed this for sale on ebay and was wondering if anybody may be able to fill me in on some specs for it
New - 300W LME49810 Mono channel Audio Amplifier Board - eBay Amplifiers, Audio, Video, Car Parts, Accessories, Cars, Bikes, Boats. (end time 15-Apr-10 05:13:34 AEST)
I would like to know what impedance it is meant to run at and what it could drive (ie lower impedances such as 2 ohms) if the supply voltage was lowered and if other circuit mods would be required to do so.
Cheers, Junglejuice
download the National datasheet.
Yeah I have that, so how using the above circuit determine what the power would be @ 4 ohms or 2 ohms?
The supply is +/- 45 VDC. For 4 Ohm, you will get around (40*40)/(2*4) = 200 W (assume min Vce = 5 V).
Ok that makes sense, on the board it has 300w listed so using the formula above for a 2 ohm load would 400w, is this too much for these transistors to handle or do I need to lower the supply voltage to 40v to give an output of 300w @ 2 ohms? 400w @ 2 ohms would be better though....
here is some extra reading on this subject.
can you please state way? is it at A class operation?
sounds like way too much dissipation..
take a pair of 130W output devices. That's a total of 260W.
Divide by 4. This results in an estimate of maximum output power of approximately 65W.
Determine what load you intend to use. Let's make that 8ohm for this example.
Determine as a first guess the supply rails for 65W into 8r0.
Vrms = sqrt( max power * load resistance) = 22.8Vac
Vpk = sqrt( max power * load resistance * 2) = 32.2Vpk.
Allow about 6V for losses and PSU sag under full power.
You need ~+-38.2Vdc to get 65W into 8r0 from one pair of 130W devices.
This should be able to drive a real 8ohm speaker reliably if normal sized transformer and normal sized smoothing capacitance and normal sized heatsink are all used.
This will not drive a 4ohm speaker. That original set of assumptions applied to an 8ohm load.
If you had 2pair of 250W devices (total 1kW) and a 4ohm load, go through the same process and determine first guess for maximum output power and PSU requirement.
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Steven S. Miller Construction builds quality custom homes in the Twin Falls area. With over 18 years of experience in the industry, Steven S. Miller Construction understands that your home is one the most important things in your life and we take pride in building your dream home.
Our customers have a hands-on role in every step of the production process. Call us today and take the first step in building your dream home!


text:
Looks like a fun forum. I've been shooting since I was 7 (more years than we'll discuss) and have recently become more regular at the range in the interests or rebuilding skills and just 'cause it's fun.
Also reload, mostly metallic stuff (45ACP, 30-06, .357, etc.) but getting into shotshell reloading as well.
Mostly carry in the mountains. I'm not looking for a trophy but I'd rather eat it than it eat me...:snipe:
Hope to learn a lot and share the fun.


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Set of 3 Rose Bushes
Cozy cotton Meow Quilt
Mock Rocks
Rolling Storage Carts
Patriotic Star Jacquard Tablecloth
Hose Splitter
Ribbon Braid Hat
Personalized Lawn/Garden Plaque - Dragonfly
Slim Slide-Out Cart
Spin Duets will dance in the slightest breeze! Love color? These spinners captivate with a sublimated design on polyester fabric. An intertwined spiral design creates mesmerizing motion with each whisper of the wind. Weather resistant fabric is attached to a fiberglass frame to maintain its sturdy shape. A heavy-duty snap swivel is included for easy hanging. Simple assembly. 15" Diam. x 25"L.
"Colorful & durable: Doesn't get any easier than this... take it out of package & hang it up. Have already gotten many compliments on the vibrant colors." - Customer in Southwick, MA
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text:
Hi to all,
I have both BT3 final & BT4 beta installed under Virtual Box and both are working great with my ALFA AWUS036H 500mW usb wireless card. As the topic of my post says I am looking for some tools that I can perform a Man-in-the-middle attack in my wireless network.
I looked on Airsnarf-ng and Airbase-ng at the moment but I couldn't understood the main differences between these two. Are both do the same things? It seems to me that Airsnarf has all the functionalities that I really need (rogue AP creation,httpd server, DNS server). These are the minimum features I need.
So if there is a chance to make my wireless card stated above to work with Airsnarf-ng then that will save me a lot of time.
I found already a tutorial of how to setup airsnarf in BT3. I followed exactly all the steps written in that but when I ran it I couldn't really understand all the error messages.
The link where I found this tutorial was
Is airsnarf-ng still used as a prefered tool for MITM attacks or are there any other tools that it will worth better looking at.
I looked in this link http://backtrack.offensive-security....Howto:Airsnarf but the reason why I didn't follow this HOW:TO
was because it was written somewhere in the forum that it wasn't a stable solution!
As last....if you recommend me to use airsnarf-ng where do you suggest me running it on BT3 or BT4b?
Thank you very much and hopefully someone can give me useful information about that!


text:
Due to the peculariaties of my biography and literary interests I have read many defenses of genres and minority art forms or literatures over the years. They all tend to fall into the same traps. Lev Grossman's opinion piece for the Wall St. Journal does a decent job of avoiding some of these traps, but still not quite as deftly as I would personally prefer.
Generally the core problem with such defenses is that they seek for respectability — literary respectability. They usually do this by:
1. Pointing out examples in their field that are unusually well-crafted and share strong similarities with literay fiction. Lev Grossman does this when he mentions George R. R. Martin and Sussana Clarke* (and Neil Gaiman). Gene Wolf's work also often gets deployed in this same way.
2. Acknowledging the field's lowbrow roots but then pointing out how the field is maturing (and usually the maturation = more works that have the marks of literary realism [see #1]).
3. Weakly throwing up hands and pointing out that people like to be entertained and read a good story and what's the harm of that.
There is some merit to each of those approaches, but they all fall down in that they, fundamentally, defer to literary fiction (or rather literary realism) and the sub-genres it has deemed acceptable (including magic realism).
Grossman tries to do better than this by pointing to the fantastic roots of fiction and by observing that all works of fiction are, well, fiction. Both good points, but also obvious ones that don't carry much weight with those that worship at the altar of literary realism or are inimical to the whole notion of narrative fiction itself. For example, Grossman writes: "When it comes to novels, fantasy is the rule, not the exception. If anything, it is realist literature that pretends to be real." Very true. But that does nothing to address the attitude that literary realism is the highest form of such pretending and that it gets closer to revealing whatever it is that we expect literature to reveal (character? truth? life? reality?).
The thing is that the proponents of literary realism** assert the superiority of the form (and you'll see a couple of examples of that in the comments section to Grossman's opinion piece), but are actually rather hard-pressed to prove it. Indeed, much of literary theory of the past 50 years seriously undermines it. And yet it in terms of critical and cultural activity (prizes, syllabi, criticism, academic appointments, anthologies, class offerings, best of lists), it remains the highbrow power in narrative fiction and other genres and minor literatures are excpected to defer to it and position themselves in relation to it. Genre writers can be gentrified by literary realists by proving their commitment to literary tropes and style. But, of course, only a few are let in to the club and, sadly, those who are the most on the edges too often feel the need to draw lines between them and their less gentrified cousins. Which then, often, leads to a petulant or resentful keening from the genres and the minor literatures. A "why not us too?"
I say stop it. Just stop it.
Don't do so by, as certain genre writers have, decrying it as elitist or morally bankrupt. Do it by showing what you can do that other genres can't (and I think that literary realism is a genre among genres and should be treated as such). Or by showing how you can do it in ways that they can't. Let me give one brief example: I would submit that a really thoughtful, honed idea sci-fi short story is of equal literary value to a really good piece of epiphanic, honed literary realism. Both do very different things with the form. But they can be equally important, artistic, entertaining, revealing — whatever you want to assign to the function of reading fiction.
Another example: I think The Road by Cormac McCarthy is a failed genre novel. I found much of the reaction to it amusing because it betrayed the literary realists lack of awareness of the field of post-apocalyptic and horror fiction***.
There's more I should say: about pulp fiction and plot vs. prose; about the recent slumming in the genres by literary authors; about the reaching for literary acclaim by genre authors; about the market and academia; about cultural signifiers and the beautiful shame of fandom. But I'll leave with this simple thought: I don't. You don't. We don't — need no literary respectability.
* Now don't get me wrong. Both A Song of Ice and Fire and Jonathan Strange & Mr. Norrell are major favs of mine.
** Now don't get me wrong. I also love literary realism. I'm one of those annoying people who loves the novels of Henry James and Tolstoy and Cather and Marilynne Robinson and has actually read Ulysses all the way through, etc.
*** Michael Chabon's review of The Road (excerpt only unless you login; it's also found in his essay collection) and his reading of it as a horror novel redeemed it somewhat in my eyes.


text:
Earl's Suspender Capo
A brand new way to capo the 5th string of your banjo. For the very first time, you can capo the 5th string at any fret without installing screws, spikes or damaging your banjo.
Easy to use Banjo Capo for playing in different Keys. Rubber pad holds the strings down, nickel plated hardware. The standard of the industry, a popular capo. Easy to remove and keep in your pocket.
The nickel plated, long 5th string sliding capo is easy to use and gives you maximum versatility in your choice of alternate tunings.
These Deering Banjo Spikes are used to capo the 5th string of your banjo. Tiny and unobtrusive these spikes will allow you to capo the 5th string of your banjo easily. The most common approach is to install spikes at frets 7, 9, and 10. They are installed by drilling a tiny hole in the finger board and carefully tapped into the hole.
Sold in packets of 12.
**If you have not installed these previously please contact Deering or a local luthier.**
email@example.com | (800) 845-7791 | 3733 Kenora Dr., Spring Valley, CA USA 91977


text:
E-Mail Hoax Beware Of Fake Microsoft Update E-mail 3 Rivers Summer Tour 2009! Ask The Help Desk What Are The Squiggly Codes I Often See When Logging Into A Secure Website? Sites Of The Month Great Sites To Check Out In July Short Tutorial Deleting Multiple E-mail Messages Using Column Headings
Welcome to eCurrents
We offer helpful information about many activities in this July issue, so no matter what's keeping you busy these days, you're covered. Are you worrying about Internet security? Don't miss the latest news about fake security alerts that pretend to be from Microsoft but are really from spammers. Are you struggling to keep up with an overflowing Inbox? Learn a fast and easy way to delete multiple e-mail messages simultaneously. Do your summer plans include baking a special cake, hiking and camping in a new place, improving the quality of your sleep, or helping out a worthy cause? If so, you'll find plenty of great ideas in Great Sites. The goal of each of our monthly eNewsletters is to keep our subscribers informed regarding their Internet connection and to improve their Internet experience. We think you'll find this information interesting. If, however, you'd prefer not to receive these bulletins on a monthly basis, please see the links at the bottom of this e-mail to manage your preferences. To see what's inside this issue, simply scroll down the eNewsletter or click on the links within the index to the left. Thanks for reading! - The eCurrents Team
E-Mail Hoax - Beware Of Fake Microsoft Update E-mail
If you receive an e-mail that appears to come from Microsoft about an Update for Microsoft Outlook/Outlook Express (KB910721), do NOT click on the link or open the attachment. These e-mails, while quite convincing in their appearance and lack of the usual spammers' misspellings, are actually fake security alerts. They are designed to get you to download Trojan malware in the form of a file named officexp-KB910721-FullFile-ENU.exe. Examples of the e-mail subject lines are:
Take a look at one such e-mail: This is a good opportunity to remind you that Microsoft never sends e-mails about a single security update. All Microsoft security updates are released together on the second Tuesday of each month and are posted at http://update.microsoft.com. You can visit this site monthly to manually upload your choice of available updates. However, Microsoft recommends that you get the updates delivered automatically to your PC. To learn how to turn on automatic updating for your particular operating system, go to: http://www.microsoft.com/windows/downloads/windowsupdate/automaticupdate.mspx. You can also sign up to receive the Microsoft Security for Home Computer Users Newsletter at http://www.microsoft.com/protect/secnews/default.mspx. In addition, don't forget to keep your antivirus software updated, and remain cautious about opening e-mail attachments (especially executable files ending in .exe).
Back to Top
3 Rivers Summer Tour 2009!
3 Rivers Communications' marketing, sales and customer service crews are hitting the road again this summer, visiting with co-op members and participating in annual local events. We'll be setting up our booth at fairs, serving up food at our customer appreciation picnic and driving our purple pick up truck in parades. Here's a list of where we'll be and when. Make sure you say hello!
Attention 3 Rivers' DIRECTV customers: Just a reminder that if you subscribe to DIRECTV through 3 Rivers, please call us directly with any customer service or technical questions at 1-800-796-4567.
Ask The Help Desk - What Are The Squiggly Codes I Often See When Logging Into A Secure Website?
Question: When I'm logging into a secure website, I often see squiggly codes such as a distorted word or phrase that I have to copy. I heard these codes are supposed to stop spammers. How do they work? Answer: You're talking about CAPTCHAs, which stands for Completely Automated Public Turing test to tell Computers and Humans Apart. These codes are alphanumeric and made of several distorted characters. Users are asked to copy the characters into an empty field—a task that's easy for a human but difficult for a computer. This is an example of a CAPTCHA:
CAPTCHAs are designed to "trip up" the automated software used by spammers. While this software is good at reading text, it has a hard time with distorted characters and will therefore be denied access to whatever website it was trying to exploit. Spammers' programs can sometimes crack a CAPTCHA, but it's a complicated process with a low success rate so most spammers are still blocked.
Sites Of The Month - Great Sites To Check Out In July
Ace Of Cakes http://foodnetwork.com/ace-of-cakes - Art and cuisine meet on the fun new Food Network show, Ace of Cakes. The show presents the creations of Chef Duff and his staff as they decorate up to 20 cakes per week, including whimsical designs and detailed replicas. The Ace of Cakes website features videos highlighting the show's sweet creations and cool cakes baked by viewers. Sleep On It http://sleepfoundation.org - The National Sleep Foundation website is packed with helpful information about sleep including articles, videos, and quizzes. You can learn about the correlation between exercise and sleep, food and sleep, how medical problems can affect sleep, and more. The site also lists international sleep care centers, and a variety of online communities to share your experiences with sleep and sleep disorders. YouTube U http://youtube.com/edu - What do Dolly Parton and President Obama have in common? They both have recently delivered university commencement addresses. You can view them, as well as other addresses and interesting lectures at the YouTube EDU site. You'll find videos on medical, scientific, political, musical, business, and other topics from YouTube's college and university partners. The Great Outdoors http://gorp.away.com - If you love to enjoy the great outdoors, you'll love this site. Find the best places to go worldwide and discover what to do once you get there. Featured activities include hiking, camping, biking, fishing, paddling, and climbing. Get advice on gear, clothing, and electronics, and tips on things like what to pack for a backpacking trip, how to find a good trail, international travel, and family trips. Reader To Reader http://readertoreader.org - Reader to Reader is a nonprofit organization dedicated to "bringing books, free of charge, to needy school libraries and public libraries across the United States." You're invited to donate books or money via their website to help make it happen. When you visit the site, you can also read about the organization's programs and success, and check out their blog to learn about recent activities.
Short Tutorial - Deleting Multiple E-mail Messages Using Column Headings
It can be a daunting task to keep up with managing your e-mail, so here's a way to save some time when you're ready to clean out your message folders. By using column headings, you can quickly delete all messages from a specified sender simultaneously—much easier than hunting down that person's messages scattered throughout your Inbox. Simply follow the steps below to delete multiple messages from a particular sender: Deleting Multiple E-mail Messages Using ... - E-mail Program: Outlook Express 6 - Operating System: Windows XP
Deleting Multiple E-mail Messages Using ... - E-mail Program: Windows Mail - Operating System: Windows Vista
Deleting Multiple E-mail Messages Using ... - E-mail Program: Mozilla Thunderbird 3.0 - Operating System: Windows XP or Vista, or Mac OS X 10.5


text:
Modern and Lovely Apartment
Size60 m²
Floor No.0
- Family/child friendly
- Kitchen
- Air conditioning
- Suitable for events
- Dishwasher
- Internet/WiFi
- Baby cot
Accommodation summary
The comfortable and modern apartment consists of a living and dining area with dining table and sofa that becomes a double bed, fully equipped kitchenette for cooking. On the ground floor there is a bathroom with a shower. The first floor is a very relaxed ambient thanks to the choice of materials with which it was designed in warm parquet floors and ceilings. You will find the main bedroom and a second bedroom with two single beds. Both rooms are extremely bright. Also on the first floor there is another bathroom with a shower and a hairdryer.
The apartment is set in an exclusive complex that consists of 6 apartments completely restructured and refurbished, transforming its outside area into an enchanting garden that can be used to relax while reading and sipping on a drink in the warm weather.
The reconstruction of the building has resulted in an environment where classical elements of the Venetian tradition are combined with elements of modern and elegant styles. Finally the residence has what in Venice is called "porta d'acqua", which, in a city whose streets are canals, is the main entrance where still today our guests can arrive or depart from with water taxis.
This apartment is set in a quiet area but is also close to shopping streets. The Cannaregio district is a well-stocked area with supermarkets, fruit and vegetable markets, bars and typical Venetian restaurants. It is just a short walk to famous sights like the Strada Nuova, il Mercato di Rialto, Saint Mark's square and the palazzo Ca' Vendramin Calergi, home to the municipal Casino of Venice without neglecting that around you can make wonderful excursions such as visits to campo del Ghetto Novo, an ancient Jewish settlement.
House rulesSecurity Deposit: €250
Arrival: between 3pm and 8pm. Departure: between 9am and 10:30pm.
Arrival from 8pm to 11pm: 20 euros. Arrival from 11pm to midnight: 40 euros. Arrival from Midnight to 3pm: during this time period, we do not allow guests to check in. (Luggage deposit places can be found at train and bus station for a fee - so please check directly with the relevant locations about the possible arrangements).
Extras that can be ordered upon request and prior to or during arrival (but please give us as much notice as possible): Baby cot: €10; change of sheets and towels (at least 24 hours' notice): €20; additional cleaning of the apartment during your stay: €50 to €70; each request for additional linen will be charged at €10 per person and will be charged to the company that you have put in charge of the reception and cleaning or to the landlord.
Tourist tax: varies from € 0.75 in low season to € 2.00 in high season. It is due only for the first 5 days. Kids under 10 years of age do not pay. Children from 10 to 16 years of age have a 50% reduction.
|Maximum Nights
|1 year
|Check-in After
|Check-in before
|On arrangement
|Check-out Before
|Cancellation Policy
Reviews for this host's other properties
We loved Venice, but sadly the accommodation let the holiday down a little, wasn't terrible, but for the money and the lovely photos didn't live up to our expectations. Was far to small and ants everywhere, probably won't book with Wimbu again, sorry.
The apartment is sober, good, clean. The pictures on the website (which is also what warning devices are not up-to-date) we expected a more spacious apartment and not facing a wall in a small alley. Therefore dark and no sun. The location itself is perfect as a base to navigate all sides of Venice; quiet and nice neighborhood again nice to stay between the Venetians. Kitchen can be better furnished: no utensils to join stir in pans no colander, no garlic press, no cheese grater, only one large pan ... that does not fit right in Italy. Moreover, the corkscrew and toaster was broken. All in all we found the price-quality ratio is not optimal ...
Het appartement is sober, goed, schoon. Door de foto's op de website (die ook wat inrichting betreft niet up-to-date zijn) verwachtten wij 'n ruimer appartement en niet met uitzicht op 'n muur in 'n klein steegje. Daardoor donker en geen zon.
De locatie op zich is perfect als uitvalsbasis om alle kanten van Venetie te bevaren; rustige leuke buurt en ook weer leuk om tussen de Venetianen te vertoeven.
Keuken kan beter ingericht worden: geen gerei om mee in pannen te roeren, geen vergiet, geen knoflook pers, geen kaasrasp, alleen 1 grote pan...dat past toch niet bij Italie. Bovendien was de kurkentrekker en broodrooster kapot. Al met al vonden wij de prijs-kwaliteit verhouding niet optimaal...
We had a great time in this apartment - it is very well located, close to the market and shops. After the three-storey walk, the apartment is big enough and functional. The master bedroom is large and a bedroom with two single beds. The last bed is on a mezzanine above the kitchen which is not ideal, but it has a nice view of Venice! Therefore recommend!
Nous avons passé un super moment dans cet apartement - il est tres bien situé, proche du marché et des commerces. Apres les 3 étages a pied, l'appartement est assez grand et fonctionnel. La chambre parentale est grande et une chambre avec 2 lits simples. Le dernier lit est sur un mezzanine au dessus de la cuisine qui n'est pas ideal, mais qui a une vue sympa sur Venise!
A recommander donc!
The apartment was beautifully decorated and with good facilities. Location was also as expected with a small courtyard at the back. Very good arrangements on arrival so we were accompanied to the apartment (addresses not much good in Venice unless you know where to go!) and told where the nearest supermarket was. Local restaurants and cafes have a really "local" ambience even though you are only 10 minutes walk from Rialto. Winter was fabulous-glorious blue skies every day-and plenty of bustle. New Year exciting in St Mark's Square- we wonder how we'll ever top this for 2014 !!! Heartily recommend !
Very nice apartment. Couldn't log in into the wifi, but everything was fine.
The apartment was just as described and matched the pictures. It was close enough to all the tourist hot spots without having that crowded feeling. The famous fish market and Rialto bridge are all within a 5 minute walk. I would definitely book again with this host.
Very pleasant stay, even in January the lights are beautiful. the contact person for the keys to our arrival c is very good password. the geographical situation of the apartment is very good, quiet close to a supermarket and a bar with very pleasant every Thursday evening a group of free jazz. since your arrival in Rome Piazzale bus you can go on foot up a meeting point Riva di Biasio vaporetto you save 7 euros per person. It takes 10 minutes walk without getting lost haha!
tres agreable sejour ,meme en janvier les lumieres sont magnifiques.
le contact de la personne pour les cles a notre arrivee c est tres bien passe.
la situation geographique de l appartement est tres bien, au calme pres d un supermarche et d un bar tres agreable avec tout les jeudis soir un groupe de jazz gratuit.
depuis votre arrivee en bus a piazzale Rome vous pouvez aller a pied jusqu a un point de rencontre riva di Biasio du vaporetto vous economiserez 7 euros par personne.
il faut 10 mn a pied sans se perdre haha!!!!
Beautiful house, clean and tidy very convenient as the location (very close to Rialto and Piazza San Marco). Equipped with all the comforts, from TV to heating! The only thing I would suggest is to change the mattress sofa bed because it is very uncomfortable but otherwise everything was perfect for a vacation in Venice is perfect!
Bellissima casa, ordinata e pulita molto comoda come posizione (vicinissima a Rialto e a Piazza San Marco). Dotata di tutti i comfort, dalla tv al riscaldamento! L'unica cosa che consiglierei è di cambiare il materasso del divano letto perchè è decisamente scomodo ma per il resto è stato tutto perfetto per una vacanza a Venezia è l'ideale!
Recommend without hesitation, a wonderful apartment in quiet residential location, away from tourist hussle yet only 10 mins walk from station and 5 mins from boats. Close to shops and restaurants. Spacious, modern, excellent value for money. Also has small courtyard. Very helpful host.
We were 5 architects visiting venice for the architecture Biennale. The appartment was next to Arsenale, and it was even more great than it apears in the advertisment photos.
Realy clean, nice, good location, cheap.
- Quality assured accommodation
- Every host is verified by telephone
- Secure payment via SSL encryption


text: METS is the world's largest exhibition of equipment, materials and systems for the international marine leisure industry. This year the show celebrates its 25th anniversary! There will be over 1,300 exhibitors from over 39 countries, over 19,000 professionals from over 96 countries and thousands of new and award winning products. For more visit www.metstrade.com.


text:
President Obama continued with his Convince America About Libya Tour on Tuesday, granting interviews to CBS, ABC and NBC to discuss U.S. intervention in the North African nation while pointing out that "each country in this region is different." —KA
The "noose has tightened" around Qaddafi as "people around him are starting to recognize that their options are limited and their days are numbered," Obama said on the "CBS Evening News," one of three interviews he gave to broadcast networks yesterday.
"We've got to ratchet up our diplomatic and our political pressure on him," Obama said. "So that at some point he makes a decision to leave."
Obama's network interviews follow a March 28 nationally televised address in which he said the U.S. committed its armed forces to a United Nations-backed mission in Libya to protect civilians from troops loyal to Qaddafi, who has been battling opposition groups for control of the country.
In the interviews with the news shows on ABC and NBC, along with CBS, Obama said the criteria he outlined for military involvement in Libya cannot be compared with the situation in Syria, another site of anti-government protests, or elsewhere in a region that has been swept by anti-government protests.
White House / Pete Souza
President Obama holds forth about Libya at the National Defense University on Monday.


text:
Submit News
Cooler Master Airs Mechanical Keyboards, New Cases, and More
Jason Mick
January 15, 2013 8:04 PM
Recipient E-mail
Sender E-mail
Component maker has exclusive contract on desirable "green" CHERRY Switches
Wrapping up one of the odds and ends of what we saw at the
2013 Consumer Electronics Show
, we spent some time talking with
Cooler Master, a top Taiwanese manufacturer
, of computer components and peripherals.
Cooler Master is among the component makers to jump on the revival of mechanical keyboards. It is offering a revamped series of keyboards this spring, with a broad range of responses.
Mechanical keyboards share a common component for the key-switches -- so called "CHERRY Switches" -- special switches manufactured by a subsidiary of German automotive electronics giant ZF Friedrichshafen AG. Each kind of CHERRY switch has a different response and goes by a different color.
Cooler Master has one of the broadest lines of keyboards in terms of different colors of CHERRY Switches. It is offering a brown CHERRY, which offers a softer keystroke with "more bump". It's also offering the very loose/clicky blue CHERRY Switch. And Cooler Master has an exclusive contract to manufacture devices based on the green CHERRY Switch, a switch that offers a very nice feel that's akin to a more controlled blue CHERRY.
Cooler Master has exclusive rights to the "green" kind of CHERRY Switch.
The company is pushing out a line of stands for Apple, Inc. (
) products, supporting everything from the iPad to MacBook Pro laptops.
The company also showed off a new case, dubbed the ATCS 710 features a beefy 240 mm fan at its top and accomodates a full ATX board. The ATCS 710 ships in Q2 at an undetermined price point.
The ATCS 710 injects itself into the ATX budget-to-midrange case competition.
The reps also showed off an interesting hybrid closed-loop cooler dubbed "Eisberg". Designed and engineered in Germany,. A closed loop design, the unusual aspect is that the copper block also supports integration in custom cooling solutions via its g1/4 fittings. In other words, it's an out of the box solution that can also be expanded on for later DIY projects.
It will be sold in one variation as a block only, or alternatively packaged with a full 120 mm fan/radiator extension. It will debut in Q2 in Europe, but will jump to the U.S. in Q3.
[All Images © Jason Mick and DailyTech LLC]
"This week I got an iPhone. This weekend I got four chargers so I can keep it charged everywhere I go and a land line so I can actually make phone calls." -- Facebook CEO Mark Zuckerberg
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POUGHKEEPSIE, N.Y. – The Marist Athletics Department announced the 2013-14 men's and women's basketball promotions schedule. Over the course of the season, Marist will host 26 men's and women's basketball games at McCann Arena. For a complete promotions schedule, CLICK HERE.
Lids Team Sports will sponsor a schedule magnet giveaway on the women's and men's basketball home openers on Friday, Nov. 8, and Sunday, Nov. 10, respectively. The first 1,000 fans on each night will receive a 2013-14 schedule magnet complements of Lids Team Sports. The men's home opener will feature Marist and Elon on Sunday, Nov. 10, at 5 p.m.
On Friday, Nov. 8, the women's basketball team will play host to Kentucky, who finished the 2012-13 season ranked eighth in the country. The first 1,000 Marist students in attendance will receive a FREE "Feed The Fox" t-shirt.
In the men's basketball Metro Atlantic Athletic Conference (MAAC) home opener against Manhattan on Friday, Dec. 6, the first 1,000 fans will receive a FREE fan banner. On the same night, Marist will also welcome all youths who attended the Jeff Bower Basketball Camps over the summer back to McCann Arena.
The women's basketball team will welcome back all campers who attended Brian Giorgis Basketball Camps and Clinics on Saturday, Dec. 21, when the Red Foxes host Big 12 foe Oklahoma at McCann Arena at 7 p.m.
Marist will host Kids Night Out and CYO Night on Friday, Jan. 3, for its MAAC women's basketball contest against Manhattan at 7 p.m. Children who wear their CYO uniforms will receive FREE admission with a paid adult.
The men's basketball program will welcome back its alumni on Saturday, Feb. 1, when it hosts Niagara in a conference showdown. Tipoff is scheduled for 7 p.m. The women will host its alumnae day on Monday, Jan. 13, when the Red Foxes host Iona in a 2013 MAAC Championship rematch at McCann Arena.
The annual Pack the House game will take place when the Marist women host MAAC rival Fairfield on Saturday, Feb. 8 at 7 p.m. A Marist Sports Public Relations class will partner with the Marist Athletics Department in an attempt to sell out McCann Arena. In the seven previous years, Marist has totaled five sellouts and was crowned the National Pack the House Champion in 2011.
Coupled with a televised doubleheader on Friday, Feb. 14, the Marist women's basketball program will host Play4Kay day to benefit Miles of Hope Breast Cancer Foundation. The Red Foxes will host Quinnipiac at 5 p.m. on ESPNU. The men's basketball game against archrival Siena will follow.
The women's basketball team will welcome all girl scouts to McCann Arena on Saturday, Jan. 25, when the Red Foxes take on Niagara at 7 p.m. The men's basketball team will host its Boy Scouts Night one week prior, on Saturday, Jan. 18, when Marist hosts Rider in a MAAC contest.
Promotional nights will be updated throughout the season and are subject to change.
Season Tickets are still available for the 2013-14 campaign. CLICK HERE to purchase yours today. Stop by the Marist Ticket Office on Monday – Friday between 10 a.m. and 4 p.m. Single game bench seats can also be purchased online. CLICK HERE to purchase tickets men's basketball single game seats. CLICK HERE to purchase women's basketball single game tickets.
Group tickets are also available for men's and women's basketball games. Groups of 15 or more receive $3 admission to women's games, and $4 tickets for men's contests. Contact the Marist Ticket Office at 845-575-3553 for more information. A $1 fee per ticket will be added to each order.


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The 7-Iron decks are stamped from 7-gauge steel which requires an incredible 4,000,000 pounds of force to stamp the shell. Stamped decks minimize the number of welds and provide a smooth underside, with fewer sharp corners where material could build. These stamped decks have rounded corners which reduces the potential damage if a deck should contact trees, rocks, curbs or other fixed objects.
7-Iron Deck
See why the 7-Iron Deck is the strongest, most durable mowing deck in the industry.
The John Deere Mulch On Demand deck lets you switch between mulching and side-discharging without ever leaving your seat. Save time and money by keeping clippings off of sidewalks and driveways and out of mulch beds.
Discover Mulch On Demand™
Switch between mulching and side discharging without ever leaving your seat.


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Novelist J. Lynn led our Self-Published Bestsellers List this week, topping charts at Amazon, Apple and Barnes & Noble.
To help GalleyCat readers discover self-published authors, we compile weekly lists of the top eBooks in four major marketplaces for self-published digital books: Amazon, B&N, Apple iBookstore and Smashwords. You can read all the lists below, complete with links to each book.
If you are an author, check out our new online course–finish your book with the help of bestselling independent authors.
New Career Opportunities Daily: The best jobs in media.Add a Comment


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Product Description
Rocket Drip Tray Grate is compatible with either the Giotto or Cellini style drip trays. While the exterior dimensions of those two models are different, the insert is the same size on either model. Fits all current Rocket models and measures 10.125 inches by 5.375 inches.
Additional Information
|Rocket Espresso
|Model Number
|Product Weight (lbs.)


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Fill out the form below and we'll be in touch to arrange a free consultation, or give us a call today at 01932 868911 and we'll get back to you right away and discuss how we can help you
SIRS Europe – Leading international investigation service. Agents from intelligence, police and military backgrounds providing unrivalled experience in a host of specialist investigation services. Our business is operational twenty-four hours a day, 365 days a year. You can read more …Read More


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Article Preview
All About Alliums
By IRA CAPLAN ();
May 25, 1969,
, Section ARTS AND LIESURE, Page D53, Column , words
PROBABLY because of their relationship to the odiferous chives, leeks, garlic, shallots and onions, alliums have been out of favor for some years. But the plants are beginning to be "discovered" and for good reasons. They are astonishingly beautiful in bloom.
May 25, 1969
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Tip: Put a + in front of words that must be included. To search for Monster Hunter and only receive results with both words, use +Monster +Hunter
You look great !!
Thank you! : ) We still don´t have the official photoshoot , for now I just could upload some from the con


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George Bush misses Air Force One -- and that's all. Anti-Count My Vote effort advances. GOP could pick two convention sites to compete with each other.
Happy Friday. Former President George W. Bush says there isn't a lot he misses about being president -- well, except for one thing: Air Force One: "A lot of people ask me, do I miss much about being president? The answer's really no. I mean I miss people I served with. I miss Air Force One. In eight years, they never lost my baggage." [
Topping the news:
The Senate moved toward a final vote on Sen. Curt Bramble's bill that would nullify the Count My Vote initiative. [
] [
] Paul Rolly says this is not the first time Bramble has meddled with a proposed ballot initiative. [
-> A group of Democratic lawyers want the feds to reopen the their investigation into former AGs John Swallow and Mark Shurtleff. [
-> The GOP's site selection committee, headed by Utah's Enid Mickelsen, may pick two finalist for the 2016 convention and have the cities compete against each other during final negotiations. [
Tweets of the day:
: "Dear US, Sorry. Love, Canada"
Happy Birthday:
To former state Rep. Laura Black and Sen. Orrin Hatch's state director, Melanie Bowen, and on Sunday to former state Rep. Craig Frank and former Utah GOP executive director Ivan DuBois.
On the Hill:
Proposed legislation would allow restaurants to post a written warning saying "Notice: This establishment dispenses and serves alcoholic products in public view" instead of having the Zion Curtain. [
-> A Senate committee passed legislation raising the smoking age to 21. [
-> The House Republican caucus voted to support a resolution in favor of moving the Utah state prison. AND, be sure to check out the Trib's list of lawmakers with real estate and development ties. [
-> Rep. Jerry Anderson, R-Price, discusses comments he made that plants in some areas of the state are struggling and need more carbon dioxide in the air. [
-> Pat Bagley explores the type of gas surfacing at the Legislature. [
-> Rep. Jim Matheson proposed a combined primary election in order to reduce party polarization. [
-> Matheson talks bout his future plans, Count My Vote and Medicaid expansion. [
-> A bill allowing UDOT to raise the speed limit to 80 mph in places the agency feels appropriate is only one vote away from the governor's desk. [
-> The House passed legislation preventing local governments from banning certain breeds of dogs. [
-> Bob Bernick and Bryan Schott discuss the fight over Medicaid expansion, the Legislature's responses to Count My Vote and what happens with new revenue numbers coming next week. [
-> Proposed legislation would make Utah the first state to regulate e-cigarettes. [
-> The Senate moved a bill aimed to help teachers and school faculty intervene if they believe a student intends to commit suicide. [
-> A Senate committee advanced a bill giving enrollment priority in charter schools to children whose grandparents helped launch the school. Similar priority is already given to children whose parents founded or serve on an operating board of a school. [
-> Lawmakers passed a bill the create Utah Jazz license plates to raise money for charity. [
-> Jim Dabakis and Thomas Wright give their takes on some of the biggest issues of the week. [
-> Gov. Gary Herbert addresses whether there is a need for a statewide anti-discrimination law. [
-> Herbert also talked about defending the anti-gay marriage Amendment 3. [
-> The Rally for Recovery stressed the importance of treating mental illness and substance abuse disorders and called for the state to accept a full Medicaid expansion. [
In other news:
Businessman Bob Fuehr discusses challenging Mia love for the GOP nomination for the 4th Congressional District. [


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Cadillac Dealership Reviews Discussion, Paul Masse Cadillac-Woonsockett, RI in Cadillac Reviews; After trying unsuccessfully to find the 08 CTS in the colors/options that I was looking for in Massachusetts over a ...
Paul Masse Cadillac-Woonsockett, RI
After trying unsuccessfully to find the 08 CTS in the colors/options that I was looking for in Massachusetts over a period of 3 weeks, I finally did my own research. I found *one* car that fit my needs, at Paul Masse Cadillac. Its about 50 miles from home, so not the most convenient place, but they were very responsive, accepted Supplier discount, and did the deal over the phone. I drove down that night and signed the paperwork, no surprises when I got there.
When I went and picked up my new CTS, however, there were serious swirls in the paint that I felt were unacceptable. After being through the wringer already looking for my car, and knowing that there were NO other ones out there like this in the Northeast, I ended up accepting delivery. After some slight pushback, I ended up speaking with the sales manager, and he nicely agreed to pick up the bill for a detail from my own guy that I use. So I left happy. Overall, a good experience, and I would recommend them.


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Atletico Mineiro overturned a 2-0 first-leg deficit on Wednesday to defeat Newell's Old Boys on penalties to reach the Copa Libertadores final, where it will play Paraguayan giant Olimpia in another two-leg series. The first leg of the final series will be played in Asuncion next Wednesday with the return leg in Belo Horizonte the following week.
Goals from
Bernard and Guilherme evened the score on aggregate at 2-2, bringing the series to overtime and eventually penalties. Ronaldinho sliced the
Newell's defense open with a superb through ball that was duly dispatched by Bernard in the third minute. However, as Newell's settled into the game, a second goal for Mineiro looked
unlikely until substitute Guilherme met Diego Mateo's weak clearance with a powerful right-footed strike from 20 yards five minutes into stoppage time in the second
Atletico goalkeeper Victor was the hero during the shootout, saving Maxi Rodriguez's effort with the score tied 2-2 after four kickers
each. Victor thanked Atletico's backroom staff for helping him analyze Newell's shootout win against Boca Juniors in the quarterfinal. "I watched those shots against Boca and I have to
thank the data staff who gave me the necessary information," he said, adding: "We've become stronger as the campaign went on. We are stronger because of what we've done so far this year. We've played
a great Copa Libertadores this year. We are focused on the final." Read the original story...


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Kathleen Van Deusen and Yixin Zhang are the winners of the 2008 Lake Oswego Farmers' Market Poster Contest in the adult and youth categories, respectively.
This is the eighth year of the popular contest, which is sponsored by the Lake Oswego Parks and Recreation Department.
More than 90 entrants from Lake Oswego participated by constructing posters representing the spirit of the market in any medium. The entries receiving the greatest number of votes in the adult and youth categories were recognized for their efforts. Winning artwork will be used for Lake Oswego Farmers' Market apparel, advertisements and signage.
The 2008 Farmers' Market will again take place in Millennium Plaza Park, running May 17 through Oct. 11. Market hours are 8:30 a.m. to 1:30 p.m.
All posters will remain on display at both Lake Oswego City Hall and the West End Building through May.
For more information about the Farmers' Market or volunteer opportunities, contact Market Manager Maddie Hillwig at 503-697-6590 or mhillwigci.oswego.or.us .
Hillwig reminded all youth that participated in the contest to stop by the Parks and Recreation Department located at the West End Building, 4101 Kruse Way, to pick up a certificate.


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Justin Bieber Could Face Felony Charges in Egging Case
As police continue to investigate Justin Bieber's egging case from last month, investigators in Los Angeles presented new evidence that could lead to a felony charge.
Per a report from the Associated Press, the district attorney's office is now determine whether or not to move forward with a felony vandalism charge against the 20-year-old singer. The crime was allegedly committed against Bieber's neighbor and caused thousands of dollars in damages.
In January, police searched Justin's home for surveillance that could lead to an arrest, and ended up finding drugs. As a result, Lil Za was taken into custody for alleged possession of cocaine. The charges were changed to felony possession of Ecstasy and oxycodone, which if the rapper is convicted, he could face to up to nine years in prison.
During the raid, Lt. David Thompson said, "I get that the eggs don't seem that significant, but it does rise to the level of a felony. There is a victim in this case who has had extensive damage done to their home. That's a serious incident."
In addition, Bieber is set to appear in court on February 14 for his drunk driving, resisting arrest and expired license charges in Miami.
Stay linked to GossipCenter for the latest on Bieber's legal woes.
Photo Credit: FilmMagic


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Selected Community Events
Go Back
http://www.wifr.com/content/events/227644051.html | 227644051
Guitars for Vets Flapjack Fundraiser
Event Dates and Times
8:00-10:00 a.m.
Applebee's Neighborhood Grill & Bar, 1675 E. Riverside Blvd., Rockford, IL 61114
Tickets cost $5 and will be available at the door. All proceeds will benefit the Rockford Chapter of Guitars for Vets. Live Entertainment
Trish Rooney - 815-985-0143
Go Back


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1. Video of Tim Keller speaking apologetically. Keller's talk is not to be missed and can be used both to edify and instruct Christians. (HT: Justin Taylor) 2. Helpful discussion of how to tackle the sometimes thorny subject of morning devotions. Covenant Life members CJ Mahaney, former pastor, and Jeff Purswell, pastor and theologian, are [Read More…]
The Week-est Link, March 21, 2008: Devotions?
March 21, 2008 by 1 Comment


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Upstream / Downstream
Explore pathways related to this product.
Research More. Spend Less.
Spend $650 or more and get 20% off*
(*Offer valid in the US only. Expires June 30, 2017)
To Purchase # 12619S
|100 µl (10 western blots)
Find answers on our FAQs page.
PhosphoSitePlus® Resource
- Additional protein information
- Analytical tools
CTR9 (D1Z4F) Rabbit mAb #12619
Western blot analysis of extracts from various cell lines using CTR9 (D1Z4F) Rabbit mAb.Learn more about how we get our images
Gallery: CTR9 (D1Z4F) Rabbit mAb #12619
Western Blotting Protocol
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
A. Solutions and Reagents
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
- 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
- 10X Tris Buffered Saline (TBS): (#12498) To prepare 1 L 1X TBS: add 100 ml 10X to 900 ml dH2O, mix.
- 1X SDS Sample Buffer: Blue Loading Pack (#7722) or Red Loading Pack (#7723) Prepare fresh 3X reducing loading buffer by adding 1/10 volume 30X DTT to 1 volume of 3X SDS loading buffer. Dilute to 1X with dH2O.
- 10X Tris-Glycine SDS Running Buffer: (#4050) To prepare 1 L 1X running buffer: add 100 ml 10X running buffer to 900 ml dH2O, mix.
- 10X Tris-Glycine Transfer Buffer: (#12539) To prepare 1 L 1X Transfer Buffer: add 100 ml 10X Transfer Buffer to 200 ml methanol + 700 ml dH2O, mix.
- 10X Tris Buffered Saline with Tween® 20 (TBST): (#9997) To prepare 1 L 1X TBST: add 100 ml 10X TBST to 900 ml dH2O, mix.
- Nonfat Dry Milk: (#9999).
- Blocking Buffer: 1X TBST with 5% w/v nonfat dry milk; for 150 ml, add 7.5 g nonfat dry milk to 150 ml 1X TBST and mix well.
- Wash Buffer: (#9997) 1X TBST.
- Primary Antibody Dilution Buffer: 1X TBST with 5% nonfat dry milk; for 20 ml, add 1.0 g nonfat dry milk to 20 ml 1X TBST and mix well.
- Biotinylated Protein Ladder Detection Pack: (#7727).
- Prestained Protein Marker, Broad Range (11-190 kDa): (#13953).
- Blotting Membrane and Paper: (#12369) This protocol has been optimized for nitrocellulose membranes. Pore size 0.2 µm is generally recommended.
- Secondary Antibody Conjugated to HRP: Anti-rabbit IgG, HRP-linked Antibody (#7074).
- Detection Reagent: SignalFire™ ECL Reagent (#6883).
B. Protein Blotting
A general protocol for sample preparation.
- Treat cells by adding fresh media containing regulator for desired time.
- Aspirate media from cultures; wash cells with 1X PBS; aspirate.
- Lyse cells by adding 1X SDS sample buffer (100 µl per well of 6-well plate or 500 µl for a 10 cm diameter plate). Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. Keep on ice.
- Sonicate for 10–15 sec to complete cell lysis and shear DNA (to reduce sample viscosity).
- Heat a 20 µl sample to 95–100°C for 5 min; cool on ice.
- Microcentrifuge for 5 min.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#13953, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
- Electrotransfer to nitrocellulose membrane (#12369).
C. Membrane Blocking and Antibody Incubations
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
I. Membrane Blocking
- (Optional) After transfer, wash nitrocellulose membrane with 25 ml TBS for 5 min at room temperature.
- Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature.
- Wash three times for 5 min each with 15 ml of TBST.
II. Primary Antibody Incubation
- Incubate membrane and primary antibody (at the appropriate dilution and diluent as recommended in the product datasheet) in 10 ml primary antibody dilution buffer with gentle agitation overnight at 4°C.
- Wash three times for 5 min each with 15 ml of TBST.
- Incubate membrane with Anti-rabbit IgG, HRP-linked Antibody (#7074 at 1:2000) and Anti-biotin, HRP-linked Antibody (#7075 at 1:1000–1:3000) to detect biotinylated protein markers in 10 ml of blocking buffer with gentle agitation for 1 hr at room temperature.
- Wash three times for 5 min each with 15 ml of TBST.
- Proceed with detection (Section D).
D. Detection of Proteins
Directions for Use:
- Wash membrane-bound HRP (antibody conjugate) three times for 5 minutes in TBST.
- Prepare 1X SignalFire™ ECL Reagent (#6883)by diluting one part 2X Reagent A and one part 2X Reagent B (e.g. for 10 ml, add 5 ml Reagent A and 5 ml Reagent B). Mix well.
- Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains wet), wrap in plastic and expose to X-ray film.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Western Blot Reprobing Protocol
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
A. Solutions and Reagents
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
- Wash Buffer: Tris Buffered Saline with Tween® 20 (TBST-10X) (#9997)
- Stripping Buffer: To prepare 100 ml, mix 6.25 ml of 1M Tris-HCl pH 6.8, 10 ml of 20% SDS and 700 μl β-mercaptoethanol. Bring to 100 ml with deionized H20. Make buffer fresh just prior to use.
B. Protocol
- After film exposure, wash membrane four times for 5 min each in TBST. Best results are obtained if the membrane is not allowed to dry.
- Incubate membrane for 30 min at 50°C in stripping buffer (with slight agitation).
- Wash membrane six times for 5 min each in TBST.
- (Optional) To assure that the original signal is removed, wash membrane twice for 5 min each with 10 ml of TBST. Incubate membrane with LumiGLO® with gentle agitation for 1 min at room temperature. Drain membrane of excess developing solution. Do not let dry. Wrap in plastic wrap and expose to x-ray film.
- Wash membrane again four times for 5 min each in TBST.
- The membrane is now ready to reuse. Start detection at the "Membrane Blocking and Antibody Incubations" step in the Western Immunoblotting Protocol.
posted June 2005
revised June 2016
Immunoprecipitation for Native Proteins
This protocol is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity.
A. Solutions and Reagents
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
- 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L of 1X PBS, add 50 ml 20X PBS to 950 ml dH2O, mix.
10X Cell Lysis Buffer: (#9803) To prepare 10 ml of 1X cell lysis buffer, add 1 ml cell lysis buffer to 9 ml dH2O, mix.
NOTE: Add 1 mM PMSF (#8553) immediately prior to use.
- 3X SDS Sample Buffer: Blue Loading Pack (#7722) or Red Loading Pack (#7723) Prepare fresh 3X reducing loading buffer by adding 1/10 volume 30X DTT to 1 volume of 3X SDS loading buffer.
- Protein A Magnetic Beads: Use Protein A (#8687) for rabbit IgG immunoprecipitation.
- 6-Tube Magnetic Separation Rack: (#7017).
- 10X Kinase Buffer (for kinase assays): (#9802) To Prepare 1 ml of 1X kinase buffer, add 100 µl 10X kinase buffer to 900 µl dH2O, mix.
- ATP (10 mM) (for kinase assays): (#9804) To prepare 0.5 ml of ATP (200 µM), add 10 µl ATP (10 mM) to 490 µl 1X kinase buffer.
B. Preparing Cell Lysates
- Aspirate media. Treat cells by adding fresh media containing regulator for desired time.
- To harvest cells under nondenaturing conditions, remove media and rinse cells once with ice-cold 1X PBS.
- Remove PBS and add 0.5 ml ice-cold 1X cell lysis buffer to each plate (10 cm) and incubate on ice for 5 min.
- Scrape cells off the plate and transfer to microcentrifuge tubes. Keep on ice.
- Sonicate on ice three times for 5 sec each.
- Microcentrifuge for 10 min at 4°C, 14,000 x g and transfer the supernatant to a new tube. The supernatant is the cell lysate. If necessary, lysate can be stored at -80°C.
C. Immunoprecipitation
Cell Lysate Pre-Clearing (Optional)
- Vortex to mix beads.
- Add 10-30 µl of 50% Protein A magnetic bead slurry of to 200 µl cell lysate at 1 mg/ml.
- Incubate with rotation at 4°C for 30-60 min.
- Pellet beads using magnetic separation rack. Transfer the supernatant to a fresh tube.
- Proceed to immunoprecipitation below.
- Add primary antibody (at the appropriate dilution as recommended in the product datasheet) to 200 µl cell lysate at 1 mg/ml. Incubate with rotation overnight at 4°C.
- Add protein A magnetic beads (10-30 µl of 50% bead slurry). Incubate with rotation for 10-30 min at 4°C.
- Pellet beads using magnetic separation rack. Wash pellet five times with 500 µl of 1X cell lysis buffer. Keep on ice between washes.
- Proceed to analyze by western immunoblotting or kinase activity (section D).
D. Sample Analysis
Proceed to one of the following specific set of steps.
For Analysis by Western Immunoblotting
- Resuspend the pellet with 20 µl 3X SDS sample buffer. Vortex, then microcentrifuge for 30 sec at 14,000 x g.
- Heat the sample to 95-100°C for 2-5 min and microcentrifuge for 1 min at 14,000 x g.
- Load the sample (15-30 µl) on a 4-20% gel for SDS-PAGE.
- Analyze sample by western blot (see Western Immunoblotting Protocol).
NOTE: To minimize masking caused by denatured IgG heavy chains (~50 kDa), we recommend using Mouse Anti-Rabbit IgG (Light-Chain Specific) (L57A3) mAb (#3677) or Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127). To minimize masking caused by denatured IgG light chains (~25 kDa), we recommend using Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127).
For Analysis by Kinase Assay
- Wash pellet twice with 500 µl 1X kinase buffer. Keep on ice.
- Suspend pellet in 40 µl 1X kinase buffer supplemented with 200 µM ATP and appropriate substrate.
- Incubate for 30 min at 30°C.
- Terminate reaction with 20 µl 3X SDS sample buffer. Vortex, then microcentrifuge for 30 sec.
- Transfer supernatant containing phosphorylated substrate to another tube.
- Heat the sample to 95-100°C for 2-5 min and microcentrifuge for 1 min at 14,000 x g.
- Load the sample (15-30 µl) on SDS-PAGE (4-20%).
posted December 2008
revised November 2013
|Western Blotting
CTR9 (D1Z4F) Rabbit mAb recognizes endogenous levels of total CTR9 protein.Species Reactivity: Human, Mouse, Rat Species predicted to react based on 100% sequence homology: Hamster, Chicken, Xenopus, Zebrafish, Bovine, Dog, Pig
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human CTR9 protein.
The PAF (RNA polymerase II (RNAPII) associated factor) complex was initially identified in yeast and is comprised of subunits PAF1, Leo1, Ctr9, Cdc73, RTF1 and Ski8 (1,2). The PAF complex plays an important role in transcription initiation and elongation by RNAPII by regulating the establishment of proper histone modifications such as histone H2B ubiquitination and the recruitment of the histone chaperone FACT (facilitates chromatin transcription) (3-5). The PAF complex also plays a role in mRNA processing and maturation by interacting with and recruiting the cleavage and polyadenylation specificity factor and cleavage stimulation factor complexes via the Cdc73 subunit (6,7). In addition, the Ski8 subunit of the PAF complex is part of the hSKi complex that regulates RNA surveillance, suggesting an important function of the complex in coordinating events associated with proper RNA maturation during transcription (1,5).
1. Jaehning, J.A. (2010) Biochim Biophys Acta 1799, 379-88.
2. Chaudhary, K. et al. (2007) Oncogene 26, 7499-507.
3. Pavri, R. et al. (2006) Cell 125, 703-17.
4. Rozenblatt-Rosen, O. et al. (2005) Mol Cell Biol 25, 612-20.
5. Zhu, B. et al. (2005) Genes Dev 19, 1668-73.
6. Rozenblatt-Rosen, O. et al. (2009) Proc Natl Acad Sci U S A 106, 755-60.
7. Farber, L.J. et al. (2010) Mol Carcinog 49, 215-23.
For Research Use Only. Not For Use In Diagnostic Procedures. Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.


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|Topic Review (Newest First)
|i have some close friends that i have known since my memory started working. we all grew up together. i still keep in touch with some old navy pals, but we are scattered over the country. i am very close with my family. the truth is, many people suck, but you have to put yourself out there to meet people and see what happens.
As I've gotten older, I've realized that it's so much better to have one or two close people than a handful of semi-acquaintances. Life is too short to waste time cultivating relationships that don't have meaning or value, so I've spent a lot of time learning to not push for frivolous friendships or sweat meeting new people.
My family is very close... I talk to my parents daily and my brother weekly. Other than that, I have my husband who is my best friend. He's the only person I have where I live... I live very rurally and work from home so I don't meet new people often. I have a couple of girlfriends I was very close with in college and when we get together it's like we never missed a beat, but we only get to see each other occasionally. Honestly, the people I talk to daily are my online friends and I'm very thankful for them.
Sent from Petguide.com Free App
|I have my husband and some of my immediate family are great. I have lunch date friends, but the two people I thought were my closest friends, I'm not sure anymore. I have a two year daughter, and these friends don't have kids and don't care about children, and don't care about my kid.
Originally Posted by Zeeva View Post
Every time I think of you, I wish I had met you when I lived in Chicago...
Originally Posted by llombardo View Post
I have great immediate family. I have several good friends, and lots of friends I grew up with. I'm a friendly person and make friends easily but I don't trust many so they are kept at arms length. I have mostly stuck with the same people for the last 30 years. A few have been added but not many.
|I have great immediate family. I have several good friends, and lots of friends I grew up with. I'm a friendly person and make friends easily but I don't trust many so they are kept at arms length. I have mostly stuck with the same people for the last 30 years. A few have been added but not many.
I'm an orphan. lost my Dad at 17 and my Mom--who was my rock--died when I was 29. I'm an only child with an only child--my son. But I am blessed with the family that I have created. Two of my closest childhood friends live nearby, my former neighbor and I talk almost every day and visit once or twice a year. My pseudo niece comes over once a week--as does my son.
When my husband had his cardiac arrest, so many of his friends and colleagues helped me. My neighbors brought food and gave me rides. I got closer to his family as well.
I do spend a lot of time alone and sometimes I do get lonely but as I've grown older I have realized that everyone has their own lives and interests. It does take an effort to keep my friendships up but I'm not afraid to be the first one to call.
I have an awesome family, and I'm especially close to my parents. And my boyfriend is practically a part of my family.
I also have two amazing best friends. Pretty much the only "outsiders" I socialize with. They have both been my good friends for years. I can't see myself being best friends with anybody else, and I wouldn't want to be.
Yes, my husband is basically my best friend. I also get along with my mom and have been doing more and more with her, and sometimes my aunts, cousins, and sister. I think we've all gotten closer as adults and after the death of my grandparents. We kind of took for granted meeting every Sunday at my grandparents' house and now that they are gone, we are putting in more effort to keep spending time together. I've reconnected with some of the cousins I grew up with who have moved away.
I have a few very close friends. I tend to do better with a smaller group of close friends than a huge group of acquaintances. I'm pretty outgoing. I work in customer support so I have a lot of training in communications and helping people and have been doing it for over a decade (I'm 29), but at my core I'm more of an introvert and "recharge" when I'm alone. I prefer to have a long lunch with one good friend rather than go out with 6 friends. Many of my best friends have moved away. I'm not the greatest and keeping in touch long distance but when we're able to get together, I feel like no time has past, we're still best friends.
|I know some fine people. However, I have an even finer family. I'm very fortunate. All my immediate family are great. They are more than family, they are also my friends. Between them and my husband/son, I rarely do things with non-family. I do consider some of them friends or friendly acquaintances... they're all good people. I know lots of iffy people as well .. but they're not my friends or anywhere close to my inner circle.
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