This antibody, ab5076, is expected to recognise the isoforms represented by NP_001614 and NP_116573 but not NP_004838 Please note: Although we have some very good Abreviews on mouse and pig, some customers were receiving inconsistent results on mouse samples. We have therefore removed mouse and pig and can no longer guarantee them. In addition, we recommend blocking in milk. Blocking with BSA gives high background.
IHC-Fr. Some customers have used this product in IHC-Fr however we have had consistent complaints in this application over the last year and can no longer guarantee performance in this application. If looking for a monoclonal anti-Iba1 alternative we can recommend our RabMAb ab178846. This has the same immunogen as ab5076.
Recombinant Human Iba1 protein (ab117478) can be used as a positive control in WB.
Please note: Although we have some very good Abreviews on mouse and pig, some customers were receiving inconsistent results on mouse and pig samples. We have therefore removed mouse and pig and can no longer guarantee them.
Our Abpromise guarantee covers the use of ab5076 in the following tested applications.
IHC-P Use a concentration of 2 - 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 0.3 - 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human Iba1 peptide (ab23067).
We received several excellent Abreviews on this antibody working with mouse, however, mouse is not a batch tested species and we cannot guarantee that this antibody will work on mouse. Some of our customers have observed high background in mouse samples. We recommend blocking in milk. Blocking with BSA gives high background.
Actin-binding protein that enhances membrane ruffling and RAC activation. Enhances the actin-bundling activity of LCP1. Binds calcium. Plays a role in RAC signaling and in phagocytosis. May play a role in macrophage activation and function. Promotes the proliferation of vascular smooth muscle cells and of T-lymphocytes. Enhances lymphocyte migration. Plays a role in vascular inflammation.
Detected in T-lymphocytes and peripheral blood mononuclear cells.
Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.
Paraffin embedded sections of human lung were stained for Iba1 with ab5076 at 5 μg/ml in immunohistochemical analysis.
Free floating sections of rat striatum/microglia were stained for Iba1 with ab5076 at 1/2000 dilution in immunohistochemical analysis. Rabbit Anti-Goat IgG Biotin was used as the secondary antibody at 1/200 dilution.
Lanes 1-14: Blocked in 3% milk for 1 hour (RT). Lanes 15-16: Blocked in 5% BSA for 1 hour (RT).
Lane 1: exposure 1 minute. Lanes 2-16: exposure 4 minutes. Abcam recommends blocking in milk for cleaner blots with reduced background, in comparison to BSA.
This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab5076 (anti-Iba1 antibody; 1 ug/mL) for 18 hours at 4°C.
ab5076 staining Iba1 in the Macrophage cell line Mono-Mac 6 by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 20 minutes at 24°C. Samples were incubated with primary antibody (1/500 in PBS) for 16 hours at 4°C. An Alexa Fluor® 555-conjugated Rabbit anti-goat IgG polyclonal (1/1000) was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody (ab5076)Image courtesy of an anonymous Abreview.
ab5076 staining Iba1 in rat glioblastoma cell line C6 by Immunocytochemistry/ Immunofluorescence.Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab5076 at a 1/50 dilution for 16 hours at 4°C. The secondary used was a Cy3 conjugated rabbit anti-goat polyclonal used at a 1/120 dilution. Nuclei are counterstained with DAPI.
AIF1 expression in an inflammatory foci in a skin biopsy. Paraffin embedded sections were deparaffinized and boiled in 10mM citrate buffer for 30 min in a microwave to expose the AIF1 antigen, and then blocked with 5% donkey serum for 20 min. Slides were incubated for 40 min with goat-anti-AIF1 antibody (1:100) and rinsed in three changes of PBS for 1 min each. A secondary antibody (donkey-anti-goat-FITC conjugated IgG; 1:50) was then applied to the slide for 40 min. Sections were washed again to remove the unbound antibody. The slides were counterstained with DaPI and viewed with a Nikon epi-fluorescent microscope. AIF1 positive cells appear green and the nuclei are stained blue. Numerous cells expressing the AIF1 protein are located around a vessel. (400X). Picture from Rreview by Carol Artlett submitted 30 July 2004.
ab5076 staining Iba1 in guinea pig brain tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in sodium citrate and then blocking with 3% serum was performed for 30 minutes at RT. The primary antibody was used at dilution at 1/200 and incubated with sample at 2% blocking serum for 18 hours at 4°C. A Biotin conjugated horse polyclonal to goat IgG was used undiluted as secondary antibody.