
vestten bloat normalisation

vestten bloat daattayude normalisation ennathu vividha pareekshanathmaka chikilsakalkku keezhilo tissuekalilo vikasana ghattangalilo ulla oru blottinteyo jellinteyo paathakaliludaneelam oru nirddishta proteente aapekshika samriddhiye thaarathamyam cheiyunna oru vishakalana gattamaanu. vestten bloat daattayil ninnu labhikkuna nigamanagalil vittuveezhcha cheyyaan kazhiyunna, asthiramaaya sample thayyarakkal, jell lainukaliludaneelam asamamaya sample loading allengil asamamaya protein transfer thudangiya pareekshanathmaka pishakukalile vyathiyanangalil ninnu undaakunna phalangal kuraykkuka ennathaanu normalaiseshante mothatthilulla lakshyam. nilavil, vestten bloat daatta saadhaarana nilayilaakkunnathin rund reethikalunda (i) house keeping protein normalisation, (ii) motham protein normalisation.
nadapadikramam.
jell allengil bloating membranilaanu normalisation nerittu sambhavikkunnath. aadyam, karayulla jell allengil bloat chithreekarikkukayum oro paathayilum taargett proteinu chuttum oru deerghachathuram varaykkukayum deerghachathurathinullile signal theevratha alakkukayum cheiyunnu. athe jellilo blottilo kandethiya loading aantharika niyanthranatthinte signal theevrathayumaayi bandhappett labhicha signal theevratha saadhaaranavalkkarikkaam. protein paadukal upayogikkumbol, karayude tharatthe aashrayich, rogaprathirodha parishodhanaykk munbo sheshamo thiranjeduttha qara upayogichu membron incubate cheyyaam.
house keeping protein niyanthranangal.
β-aaktin, gyapd, echprt1, rplp1 ennivayulppedeyulla house keeping jeenukalum proteinukalum palappozhum vestten blottukalil aantharika niyanthranangalayi upayogikkunnu, kaaranam avaa pareekshanangaliludaneelam ore thalangalil gadanaaparamaayi prakadippikkunnathaayi karuthappedunnu. ennirunnaalum, sameepakaala padanangal kaanikkunnath house keeping proteinukalude (echkps) aavishkaaram vividha kosha tharangalilum jaiva avasthakalilum mariyekkaam ennaanu. athinaal, shaasthreeya prasadhakarkkum dhanasahaya ejansikalkkum ippol phalangalude punarnirmmanakshamathayum krithyathayum urappaakkunnathin oro pareekshanatthinum saadhaaraneekarana niyanthranangal munbu saadhookarikkanamennu aavasyamundu.
fluorescent aantibodikal.
vestten blottukalile imej proteinukalilekku fluorescent aantibodikal upayogikkumbol, normalisation aavashyappedunnathu upayokthaav alavinte mukalilum thaazheyumulla paridhikal nirvachikkukayum oro aantijanileyum signal theevrathayum sample maas voliyavum thammilulla rekheeya bandhathe savisheshamaakkukayum venam. taargett protein normalisation niyanthranavum kandethalinte chalanathmaka paridhikkullil fluores cheyyendathundu. pala echkipiesukalum uyarnna thalatthil prakadippikkukayum uyarnna reethiyil prakadippikkunna taargett proteinukalulla upayogathinu munganana nalkukayum cheiyunnu. kuranja express cheiyunna proteinukal ore paadukalil kandethuka prayaasamaan.
fluorescent aantibodikal vaanijyaparamaayi labhyamaan, phalangalude sthiratha urappakkaan poornnamaayum savisheshathayulla aantibodikal shupaarsha cheiyunnu.
fluorescent kandethal upayogikkaathappol, loading kantrol protein thaalpparyamulla protein tanmatra bhaarathil ganyamaayi vyathyaasappedanam, athinaal krithyamaaya vishakalanathinayi avaye jell electrophoresis kondu vendathra verthirikkunnu.
membran stripping.
ore paadile onniladhikam protein lakshyangal kandethumbol oru puthiya koottam dittakshan aantibodikal upayogichu membranukal neekkam cheyyukayum veendum parisodhikkukayum cheyyendathundu. phalapradamallaatha stripping taargett proteinil ninnulla durbalamaaya signalinu kaaranamakum. aantijan nashtappedunnathu thadayaan, oru membranil moonnu stripping incubationukal maathrame shupaarsha cheiyunnulloo. valareyadhikam adangiyirikkunna proteinukalil ninnulla signal poornnamaayum illaathaakkunnathu budhimuttaayirikkum, athinaal kuranja reethiyil prakadippikkunna proteinukal aadyam kandethanaanu shupaarsha cheiyunnathu.
exogenous spike-inn niyanthranangal.
echkipi alavu tissuekalkkidayil asthiramaayirikkaamennathil, antibody rekheeya paridhikkullil ariyappedunna saandrathayude shudhavum exogenous protein spike cheythukondu shaasthranjarkku thaalpparyamulla protein niyantrikkaan kazhiyum. echkipiyumaayi thaarathamyappeduthumbol, spike-inn niyanthranathinaayi vaividhyamaarnna proteinukal labhyamaan.
motham protein normalisation.
motham protein normalisationil (tii. pii. enn), taargett proteente samriddhi oro paathayileyum motham protein alavil normalise cheiyunnu. tii. pii. enn orotta loading niyanthranathe aashrayikkaathathinaal, niyanthranangalude moolyanirnnayavum echkps kandethunnathinu blottukal neekkam cheyyuka/veendum probbing cheyyuka enniva aavasyamilla. ithinu krithyatha (oro paathayilum motham proteente 0.1 microgram vare), chelavu-falaprapti, daatta vishwaasyatha enniva mechappeduthaan kazhiyum.
fluorescent paadukalkkum steyin free jellukalkkum jell/blottile proteinukal drishyavalkkarikkunnathinna prathyeka upakaranangal aavasyamaanu. paadukal paadine thulyamaayi moodanamennilla; madhyabhaagathekkaal kooduthal qara paadukalude arikukalilekku shekharikkaam. chithratthile ekathaanatha illaathathu thettaaya saadhaaranavalkkaranathinu kaaranamakum.
pree-antibody paadukal.
aantibodikal cherkkunnathinu munbu ponsiyo, koomasi, briliyant blue thudangiya ionic chaayangalum sipro rubie, deep purple thudangiya fluorescent chaayangalum upayogikkunnu, kaaranam avaa thaazhekkulla rogaprathirodhasheshiye baadikkunnilla.
proteinukalkku chuvapp pink niratthil karayidunna negatteevu charjju cheytha reversible dye aanu ponsos. ponsoyude karayude theevratha kaalakramena vegathil kurayunnu, athinaal documentation vegathil nadatthanam. valare samayathe aasrayikkunna karayude theevrathayum kuranja signal-tuu-noys anupaathavum kaaranam mosham punarnirmmanakshamathayulla ponsiyoykku 140 microgram vare oru rekheeya shreni report cheyyappedunnu.
sipro rubie polulla fluorescent chaayangalkk vishalamaaya rekheeya shreniyundu, avaa ionic chaayangalekkaal sensitiivu aanu. avaa sthiravum photostable kalarppukalum oru staanderd yuvi allengil blue-laitt traansiluminettar allengil lazer scan upayogichu drishyavalkarikkaavunnathu. charge-cuppled device camera upayogichu membranukal philim allengil digitalaayi rekhappeduthaam. sipro rubie bloat staining samayamedukkunnathaanu, koodaathe oru pathayil 50 microgram proteinil kooduthal poorithamaakum.
poost-antibody paadukal.
ponsoyekkaal sensitiivu aaya sthiramaaya poost-antibody ionic karayaanu amido black. rogaprathirodhasheshamulla roganirnayathinu sheshamaan eee qara prayogikkunnathu.
karayillaatha saankethikavidya.
steyin free saankethikavidya imajinginaayi oru inn-jell rasathanthram upayogikkunnu. eee raasapravarthanam protein kaimattattheyo thaazhekkulla antibody bandhatheyo baadikkilla. koodaathe, ithil qara/nirnnayaka gattangal ulppedunnilla, koodaathe baandukalude theevratha kaalakramena sthiramaayi thudarunnu.
tryptophan avashishtangal adangiyittillaatha proteinukal kandethaan steyin free saankethikavidyaykku kazhiyilla. kandethal praapthamaakkunnathin kuranjathu rund tryptophaanukal aavasyamaanu. 18 kinarukalkku oru pathayil 80 microgram protein 12 kinarukalkku idatharam valippamulla jellukalkku oru pathayil 100 microgram proteinumaanu steyin free normalaiseshante rekheeya paridhi. eee shreni kvaanditteevu vestten blottukalile saadhaarana protein lodukalumaayi poruthappedunnu, koodaathe vishalamaaya protein-loading shreniyil loading niyanthrana kanakkukoottalukal prapthamaakkunnu. kooduthal kaaryakshamamaya karayillaatha reethiyum aduthide labhyamaayittund. uyarnna protein lodukal upayogikkumbol, karakalillaattha saankethikavidya karakalekkaal valia vijayam theliyichittundu.

