Anti-CD31 antibody [EPR3094] is a rabbit recombinant monoclonal antibody that is used to detect CD31 in ICC/IF, IHC-P, Western blot. Suitable for Human samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Cited in over 70 publications
- Trusted since 2009
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IHC-P | ICC/IF | IP | Flow Cyt | WB | |
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Human | Tested | Tested | Not recommended | Not recommended | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info 1/100 - 1/500 | Notes We suggest to program the pressure cooker to run for 30 seconds at 125° C, followed by 10 seconds at 90° C. Then let the slide cool down to room temperature (10 - 20 minutes). The ideal fixation time will depend on the size of the tissue block and the type of tissue, but fixation between 18–24h is suitable for most samples. Positive Control: Hu tonsil tissue Perform heat-mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
Species | Dilution info | Notes | |
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Species Human | Dilution info 1/500 | Notes
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Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
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Species Human | Dilution info 1/5000 - 1/20000 | Notes Treat samples with PNGase F or phosphatase to confirm the specificity of bands if necessary. The observed band size of CD31 may not the same as predicted MWs in WB due to the different forms and modifications of CD31. Hu Isoform 1-6: 79-83 kDa (predicted) Observed band size is around 130 kDa Positive Control: HUVEC and Jurkat cell lysates (ab7899); Human spleen and kidney tissue lysate. Negative Control: NIH/3T3 whole cell lysate (ab7179). |
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Cell adhesion molecule which is required for leukocyte transendothelial migration (TEM) under most inflammatory conditions (PubMed:17580308, PubMed:19342684). Tyr-690 plays a critical role in TEM and is required for efficient trafficking of PECAM1 to and from the lateral border recycling compartment (LBRC) and is also essential for the LBRC membrane to be targeted around migrating leukocytes (PubMed:19342684). Trans-homophilic interaction may play a role in endothelial cell-cell adhesion via cell junctions (PubMed:27958302). Heterophilic interaction with CD177 plays a role in transendothelial migration of neutrophils (PubMed:17580308). Homophilic ligation of PECAM1 prevents macrophage-mediated phagocytosis of neighboring viable leukocytes by transmitting a detachment signal (PubMed:12110892). Promotes macrophage-mediated phagocytosis of apoptotic leukocytes by tethering them to the phagocytic cells; PECAM1-mediated detachment signal appears to be disabled in apoptotic leukocytes (PubMed:12110892). Modulates bradykinin receptor BDKRB2 activation (PubMed:18672896). Regulates bradykinin- and hyperosmotic shock-induced ERK1/2 activation in endothelial cells (PubMed:18672896). Induces susceptibility to atherosclerosis (By similarity). Isoform Delta15. Does not protect against apoptosis.
CD31, Platelet endothelial cell adhesion molecule, PECAM-1, EndoCAM, GPIIA', PECA1, PECAM1
Anti-CD31 antibody [EPR3094] is a rabbit recombinant monoclonal antibody that is used to detect CD31 in ICC/IF, IHC-P, Western blot. Suitable for Human samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Cited in over 70 publications
- Trusted since 2009
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Anti-CD31 antibody [EPR3094] (ab76533) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ICC/IF, IHC-P, WB in human samples.
Anti-CD31 antibody [EPR3094] (ab76533) has been cited over 80 times in peer reviewed journals and is trusted by the scientific community.
Abcam's high quality manufacturing and validation processes ensure Anti-CD31 antibody [EPR3094] (ab76533) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-CD31 antibody [EPR3094] (ab76533) has 11 independent reviews from customers.
Anti-CD31 antibody [EPR3094] (ab76533) specifically detects CD31 (UniProt ID: P16284; Molecular weight: 80kDa) and is sold in 100 µL and 1 mL selling sizes.
Conjugation-ready, carrier free format available for antibody clone EPR3094 - Anti-CD31 antibody [EPR3094] - BSA and Azide free ab207090.
Antibody clone EPR3094 is also available pre-conjugated to a variety of labels for your convenience - Biotin, Alexa Fluor® 647, Alexa Fluor® 488, Alexa Fluor® 594, Alexa Fluor® 555 (Biotin Anti-CD31 antibody [EPR3094] ab199734, Alexa Fluor® 647 Anti-CD31 antibody [EPR3094] ab218582, Alexa Fluor® 488 Anti-CD31 antibody [EPR3094] ab275989, Alexa Fluor® 594 Anti-CD31 antibody [EPR3094] ab277231, Alexa Fluor® 555 Anti-CD31 antibody [EPR3094] ab279331).
This antibody shows no cross-reactivity with rat and mouse samples in WB. However it can give some non specific staining on mouse smooth muscle tissues. Plesae contact our Scientific support for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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ab76533 staining CD31 in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Immunohistochemical analysis of endothelial colony forming progenitor cell plugs, staining CD31 (green) with ab76533.
Following antigen retrieval and blocking, sections were incubated with primary antibody (1/1000) overnight at 4°C. A Cy5®-conjugated anti-rabbit IgG (2 μg/ml) was used as the secondary antibody.
ab76533 staining CD31 in THP-1 (Human monocytic leukemia monocyte) cell line. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab76533 at 1/250 (4.9 μ μg/mL) overnight at +4°C. ab195889Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used as a counterstain at 1/200 dilution. DAPI (blue) was used as nuclear counterstain.
Confocal image showing membranous staining in THP-1 cells.
All lanes: Western blot - Anti-CD31 antibody [EPR3094] (ab76533) at 1/10000 dilution
All lanes: THP-1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 82 kDa
Observed band size: 125 kDa
All lanes: Western blot - Anti-CD31 antibody [EPR3094] (ab76533) at 1/20000 dilution
Lane 1: THP-1 cell lysate at 10 µg
Lane 2: Jurkat cell lysate at 10 µg
All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 82 kDa
Observed band size: 125 kDa
Immunohistochemical analysis of paraffin embedded human kidney tissue using ab76533 at a 1/250 dilution. Note positive staining of endothelial cells.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin embedded human muscle tissue using ab76533 at a 1/250 dilution. Note positive staining of endothelial cells.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
ab76533 staining CD31 in human muscle tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then blocked with 3% serum for 30 minutes at 20°C followed by incubation with the primary antibody at a 1/200 dilution for 12 hours at20°C. A Cy3®-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/200 dilution.
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Tissue Microarrays stained for "Anti-CD31 antibody [EPR3094]" using "ab76533"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab76533 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
This image was generated using Anti-CD19 antibody [SP291] - C-terminal ab227688, the same clone, but with a different buffer formulation.
Panel A: merged staining of anti-CD31 (gray; Opal™690), anti-CD3 epsilon (green; Opal™520) and anti-CD19 (red; Opal™570) on Formalin/PFA-fixed paraffin-embedded sections of human tonsil. Secondary antibody was Opal Polymer HRP Ms + Rb, nuclear counterstain was DAPI.
Panel B: anti-CD3 epsilon stained on T cells with Anti-CD3 epsilon antibody [SP7] ab16669 at 1/500 dilution
Panel C: anti-CD19 stained on B cells with Anti-CD19 antibody [SP291] - BSA and Azide free ab237772 at 1/5000 dilution
Panel D: anti-CD31 stained on endothelial cells and immune cell subsets with Anti-CD31 antibody [EPR3094] - BSA and Azide free ab207090 at 1/500 dilution
The section was incubated in three rounds of staining: in the order of Anti-CD31 antibody [EPR3094] - BSA and Azide free ab207090 and Anti-CD3 epsilon antibody [SP7] ab16669 for 30 mins, then Anti-CD19 antibody [SP291] - BSA and Azide free ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling CD31 with ab76533 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32mins.
ab76533 anti-CD31 [EPR3094] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
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