Name: Anti-CD31 antibody [EPR3094]
Brand: RabMAb
SKU: ab76533
Rating: 4 (12 reviews)

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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/IF, WB, IHC-P
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Biotin Carrier Free

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/IF, 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.
Positive control
- WB: THP-1 and Jurkat cell lysates IHC-P: Human kidney tissue and human muscle tissue ICC/IF: THP-1 cell lysate.
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
For more information see here.
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Dissociation constant (K_D)

K_D = 1.79 x 10 ^-10 M
Learn more about K_D
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

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human CD31 protein (ab155607)
Related Products
- Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

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/IF

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

(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

ICC/IF
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
1/5000 - 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
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

Application notes

Is unsuitable for Flow Cyt or IP.

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
modifications

Phosphorylated 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.
Target information above from: UniProt accession P16284 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 5175 Human
- Omim: 173445 Human
- SwissProt: P16284 Human
- Unigene: 376675 Human
- Unigene: 514412 Human
Alternative names
- Adhesion molecule antibody
- CD31 antibody
- CD31 antigen antibody
- CD31 EndoCAM antibody
- EndoCAM antibody
- FLJ34100 antibody
- FLJ58394 antibody
- GPIIA antibody
- GPIIA' antibody
- PECA1 antibody
- PECA1_HUMAN antibody
- Pecam 1 antibody
- PECAM 1 CD31 EndoCAM antibody
- PECAM antibody
- PECAM-1 antibody
- Pecam1 antibody
- Platelet and endothelial cell adhesion molecule 1 antibody
- Platelet endothelial cell adhesion molecule antibody
- Platelet/endothelial cell adhesion molecule 1 antibody
see all

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.
Immunocytochemistry/ Immunofluorescence - 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)

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
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)

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)

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.
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)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.
See Abreview
OI-RD Scanning - Anti-CD31 antibody [EPR3094] (ab76533)

Equilibrium disassociation constant (K_D) measurement to determine antibody affinity to the target antigen.

Click here to learn more about K_D
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] (ab76533)

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 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).
Anti-CD31 antibody [EPR3094] (ab76533)

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab76533? Please let us know so that we can cite the reference in this datasheet.

ab76533 has been referenced in 68 publications.

Rosa A et al. WASp controls oriented migration of endothelial cells to achieve functional vascular patterning. Development 149:N/A (2022). PubMed: 34931661
Anderson AE et al. An immunologically active, adipose-derived extracellular matrix biomaterial for soft tissue reconstruction: concept to clinical trial. NPJ Regen Med 7:6 (2022). PubMed: 35031598
Ly TTG et al. Inhibitory Effect of Etravirine, a Non-Nucleoside Reverse Transcriptase Inhibitor, via Anterior Gradient Protein 2 Homolog Degradation against Ovarian Cancer Metastasis. Int J Mol Sci 23:N/A (2022). PubMed: 35055132
Ren H et al. A bioactivated in vivo assembly nanotechnology fabricated NIR probe for small pancreatic tumor intraoperative imaging. Nat Commun 13:418 (2022). PubMed: 35058435
Li Y et al. Single-cell RNA sequencing reveals the multi-cellular ecosystem in different radiological components of pulmonary part-solid nodules. Clin Transl Med 12:e723 (2022). PubMed: 35184398

View all Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Frontal Cortex (Brain))

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris-EDTA

Permeabilization

Yes - .1% TritonX-100

Specification

Frontal Cortex (Brain)

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Daniel Kirsch

Verified customer

Submitted Nov 30 2020

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Cirrhosis Liver)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0

Permeabilization

Yes - 0.05% Tween 20

Specification

Cirrhosis Liver

Blocking step

sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 31 2019

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (MDA-MB-231 xenograft model)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: citric acid

Permeabilization

Specification

MDA-MB-231 xenograft model

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 01 2017

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Kidney)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citric acid pH6

Permeabilization

Specification

Kidney

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Carl Hobbs

Verified customer

Submitted Sep 06 2016

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate-buffer pH 6,0

Sample

Pig Tissue sections (skin)

Specification

skin

Permeabilization

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 18 2015

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6

Sample

Human Tissue sections (muscle)

Specification

muscle

Permeabilization

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 23 2013

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (muscle)

Specification

muscle

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrat-Buffer pH 6.0

Permeabilization

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 10 2011

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Dissociation constant (KD)

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (68)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 antibody [EPR3094]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 antibody [EPR3094]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 antibody [EPR3094]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 antibody [EPR3094]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 antibody [EPR3094]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 [EPR3094] antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 antibody [EPR3094]

Dissociation constant (K_D)

Post-translational
modifications