Recombinant Anti-CD31 antibody [EPR3094] (ab76533)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-267989-anti-cd31-antibody-epr3094-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3094] to CD31
- Suitable for: ICC, WB, IHC-P
- Reacts with: Human
Overview
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Product name
Anti-CD31 antibody [EPR3094]
See all CD31 primary antibodies -
Description
Rabbit monoclonal [EPR3094] to CD31 -
Host species
Rabbit -
Tested applications
Suitable for: ICC, WB, IHC-Pmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: THP-1 and Jurkat cell lysates IHC-P: Human kidney tissue and human muscle tissue ICC: THP-1 cell lysate.
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General notes
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.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Dissociation constant (KD)
KD = 1.79 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3094 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Biotin Anti-CD31 antibody [EPR3094] (ab199734)
- Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
- 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)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76533 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC | (2) |
1/500.
It is recommended to incubate cells with 0.1% Triton-X for 5 min to detect nuclear antigen. Use 0.3M glycine to quench autofluorescence caused by aldehydes. Positive Control: HUVEC cells |
| WB | (1) |
1/5000 - 1/20000. Predicted molecular weight: 83 kDa.
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). |
| IHC-P | (7) |
1/100 - 1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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 |
| Notes |
|---|
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ICC
1/500. It is recommended to incubate cells with 0.1% Triton-X for 5 min to detect nuclear antigen. Use 0.3M glycine to quench autofluorescence caused by aldehydes. Positive Control: HUVEC cells |
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WB
1/5000 - 1/20000. Predicted molecular weight: 83 kDa. 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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IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. 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 |
Target
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Function
Induces susceptibility to atherosclerosis (By similarity). Cell adhesion molecule which is required for leukocyte transendothelial migration (TEM) under most inflammatory conditions. 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. Prevents phagocyte ingestion of closely apposed viable cells by transmitting 'detachment' signals, and changes function on apoptosis, promoting tethering of dying cells to phagocytes (the encounter of a viable cell with a phagocyte via the homophilic interaction of PECAM1 on both cell surfaces leads to the viable cell's active repulsion from the phagocyte. During apoptosis, the inside-out signaling of PECAM1 is somehow disabled so that the apoptotic cell does not actively reject the phagocyte anymore. The lack of this repulsion signal together with the interaction of the eat-me signals and their respective receptors causes the attachment of the apoptotic cell to the phagocyte, thus triggering the process of engulfment). Isoform Delta15 is unable to protect against apoptosis. Modulates BDKRB2 activation. Regulates bradykinin- and hyperosmotic shock-induced ERK1/2 activation in human umbilical cord vein cells (HUVEC). -
Tissue specificity
Expressed on platelets and leukocytes and is primarily concentrated at the borders between endothelial cells. Isoform Long predominates in all tissues examined. Isoform Delta12 is detected only in trachea. Isoform Delta14-15 is only detected in lung. Isoform Delta14 is detected in all tissues examined with the strongest expression in heart. Isoform Delta15 is expressed in brain, testis, ovary, cell surface of platelets, human umbilical vein endothelial cells (HUVECs), Jurkat T-cell leukemia, human erythroleukemia (HEL) and U937 histiocytic lymphoma cell lines (at protein level). -
Sequence similarities
Contains 6 Ig-like C2-type (immunoglobulin-like) domains. -
Domain
The Ig-like C2-type domains 2 and 3 contribute to formation of the complex with BDKRB2 and in regulation of its activity. -
Post-translational
modificationsPhosphorylated on Ser and Tyr residues after cellular activation. In endothelial cells Fyn mediates mechanical-force (stretch or pull) induced tyrosine phosphorylation. -
Cellular localization
Membrane. Cell junction. Localizes to the lateral border recycling compartment (LBRC) and recycles from the LBRC to the junction in resting endothelial cells and Cell junction. Localizes to the lateral border recycling compartment (LBRC) and recycles from the LBRC to the junction in resting endothelial cells. - Information by UniProt
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Database links
- Entrez Gene: 5175 Human
- Omim: 173445 Human
- SwissProt: P16284 Human
- Unigene: 376675 Human
- Unigene: 514412 Human
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Alternative names
- Adhesion molecule antibody
- CD31 antibody
- CD31 antigen antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)
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) ab97051 was used as the secondary antibody at a dilution of 1/500.
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![Immunocytochemistry - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-453246-AntiCD31-antibody-ICC.jpg)
Immunocytochemistry - Anti-CD31 antibody [EPR3094] (ab76533)Immunocytochemistry analysis of THP-1 (Human monocytic leukemia monocyte) cell line labeling CD31 with ab76533 at 1/250 (4.9 μg/mL). ab195889 Anti-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. -
![Western blot - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-267988-anti-cd31-antibody-epr3094-western-blot.jpg)
Western blot - Anti-CD31 antibody [EPR3094] (ab76533)Anti-CD31 antibody [EPR3094] (ab76533) at 1/10000 dilution + THP-1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 83 kDa
Additional bands at: 125 kDa. We are unsure as to the identity of these extra bands.Blocking and diluting buffer: 5% NFDM /TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-92000-anti-cd31-antibody-epr3094-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)Image from Hofmann NA et al., PLoS One. 2012;7(9):e44468. doi: 10.1371/journal.pone.0044468. Epub 2012 Sep 7. Fig 3.; doi:10.1371/journal.pone.0044468; September 7, 2012, PLoS ONE 7(9): e44468.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. -
![Western blot - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-92004-anti-cd31-antibody-epr3094-western-blot.jpg)
Western blot - Anti-CD31 antibody [EPR3094] (ab76533)All lanes : Anti-CD31 antibody [EPR3094] (ab76533) at 1/20000 dilution
Lane 1 : THP-1 cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 83 kDa
Observed band size: 125 kDa why is the actual band size different from the predicted? -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-92003-anti-cd31-antibody-epr3094-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-92002-anti-cd31-antibody-epr3094-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)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.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-92001-anti-cd31-antibody-epr3094-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)Image courtesy of an anonymous Abreview.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. -
![OI-RD Scanning - Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-91999-anti-cd31-antibody-epr3094.jpg)
OI-RD Scanning - Anti-CD31 antibody [EPR3094] (ab76533)Equilibrium disassociation constant (KD) measurement to determine antibody affinity to the target antigen.
Click here to learn more about KD -
![Anti-CD31 antibody [EPR3094] (ab76533)](https://www.abcam.com/ps/products/76/ab76533/Images/ab76533-10-benefits-of-recombinant-antibodies.png)
Anti-CD31 antibody [EPR3094] (ab76533)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (36)
ab76533 has been referenced in 36 publications.
- Wang L et al. Homozygous MESP1 knock-in reporter hESCs facilitated cardiovascular cell differentiation and myocardial infarction repair. Theranostics 10:6898-6914 (2020). PubMed: 32550911
- Jiang Z et al. Inactivation of the Wnt/ß-catenin signaling pathway underlies inhibitory role of microRNA-129-5p in epithelial-mesenchymal transition and angiogenesis of prostate cancer by targeting ZIC2. Cancer Cell Int 19:271 (2019). PubMed: 31649488
- Lefley D et al. Development of clinically relevant in vivo metastasis models using human bone discs and breast cancer patient-derived xenografts. Breast Cancer Res 21:130 (2019). PubMed: 31783893
- Sato S et al. EPHB2 carried on small extracellular vesicles induces tumor angiogenesis via activation of ephrin reverse signaling. JCI Insight 4:N/A (2019). PubMed: 31661464
- Sun C et al. Meis2 represses the osteoblastic transdifferentiation of aortic valve interstitial cells through the Notch1/Twist1 pathway. Biochem Biophys Res Commun 509:455-461 (2019). PubMed: 30594396