Anti-CD31 antibody (ab124432)
Key features and details
- Rabbit polyclonal to CD31
- Suitable for: WB, IHC-P
- Reacts with: Mouse
- Isotype: IgG
Overview
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Product name
Anti-CD31 antibody
See all CD31 primary antibodies -
Description
Rabbit polyclonal to CD31 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse -
Immunogen
Synthetic peptide within Mouse CD31 aa 650 to the C-terminus conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available asab155816) -
Positive control
- WB: Mouse Heart and Spleen tissue lysates and TH2 whole cell lysate. IHC-P: Mouse Placenta tissue.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Images
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Western blot - Anti-CD31 antibody (ab124432)All lanes : Anti-CD31 antibody (ab124432) at 1 µg/ml
Lane 1 : TH2 whole cell lysate
Lane 2 : Mouse spleen
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 120-140 kDa why is the actual band size different from the predicted?This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab124432 overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) at a 1:10000 dilution for 1hr at room temperature and then imaged.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (ab124432) Binder BY et al. Lysophosphatidic acid enhances stromal cell-directed angiogenesis. PLoS One 8:e82134 (2013). Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Formalin-fixed, paraffin-embedded ischemic and normal mouse hindlimb tissue stained for CD31 using ab124432 at 1/1000 dilution. The Rabbit specific HRP/DAB detection kit (ab64261) was used as the secondary reagent. CD31 staining reveals the formation of larger intramuscular blood vessels in limbs receiving (h) both LPA and ASC, while (e) normal tissue and defects receiving (f) LPA and (g) ASC alone have fewer and smaller vessels.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (ab124432)IHC image of CD31 staining in mouse placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH6 for 20 mins. The section was then incubated with ab124432, 1µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Western blot - Anti-CD31 antibody (ab124432)All lanes : Anti-CD31 antibody (ab124432) at 1 µg/ml (Milk blocking - 3%)
Lane 1 : Heart (Mouse) Tissue Lysate
Lane 2 : Spleen (Mouse) Tissue Lysate
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?
Additional bands at: 90 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
Abcam recommends using milk as the blocking agent - 3%. CD31 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab124432 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
