Overview

  • Product name
    Anti-CD31 antibody [JC/70A]
    See all CD31 primary antibodies
  • Description
    Mouse monoclonal [JC/70A] to CD31
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human, Cynomolgus monkey
  • Immunogen

    Membrane preparation of a spleen from a patient with hairy cell leukaemia.

  • Positive control
    • WB: HUVEC, HeLa, human spleen, human kidney IHC-P: FFPE Human Tonsil normal. ICC-IF: HUVEC cells.
  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

Applications

Our Abpromise guarantee covers the use of ab9498 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
IHC-Fr Use a concentration of 1 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
WB 1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 82 kDa).
IHC-P Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • 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.
  • Information by UniProt
  • Database links
  • 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

Images

  • ab9498 staining CD31 in HUVEC cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1%PBS-Tween 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 overnight at +4°C with ab9498 at 1μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

     

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • IHC image of ab9498 staining in 10% formaldehyde fixed frozen tissue section of human lung.

    Non-specific protein-protein interactions were blocked using TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1 hour at room temperature. The section was then incubated with ab9498 (1μg/ml concentration) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then incubated with ab150119 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647)) and DAPI for 1 hour at room temperature.

    The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is derived directly from bound ab9498. 

    For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.

  • IHC image of CD31 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab9498, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Overlay histogram showing Jurkat cells stained with ab9498 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9498, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with methanol (5 min) used under the same conditions.

    Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • ab9498 stained HUVEC cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9498 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • ab9498 stained in HVEC cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab9498 at 5 µg/ml and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150080 (pseudo-colored red) and ab150117 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature

  • IHC image of CD31 staining in human spleen formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9498, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer 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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ab9498 at 1/100 staining human lymph node tissue sections by IHC-P. The tissue was paraformaldehyde fixed and a heat based antigen retrieval step was performed. The tissue was then blocked with serum and incubated with ab9498 overnight. An HRP conjugated goat antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Jiménez-Torres JA  et al. Patient-specific organotypic blood vessels as an in vitro model for anti-angiogenic drug response testing in renal cell carcinoma. EBioMedicine 42:408-419 (2019). Read more (PubMed: 30902740) »
  • Zhao C  et al. ETV2 mediates endothelial transdifferentiation of glioblastoma. Signal Transduct Target Ther 3:4 (2018). Read more (PubMed: 29527330) »
See all 64 Publications for this product

Customer reviews and Q&As

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1-10 of 22 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: cell conditioner 1
Permeabilization
No
Specification
lung
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 09 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (LIVER)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: CC1 Roche
Specification
LIVER
Fixative
Formaldehyde

Mr. Emmanuel Bouchaert

Verified customer

Submitted Oct 22 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Monkey Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA
Specification
kidney
Fixative
Formaldehyde

Dr. Li Li

Verified customer

Submitted Sep 19 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Rabbit Tissue sections (Heart)
Permeabilization
No
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 14 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Saphenous Vein)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
Yes - 0.3% Hydrogen Peroxide
Specification
Saphenous Vein
Blocking step
Casein in PBS as blocking agent for 10 minute(s) · Concentration: 0.25% · Temperature: 21°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 29 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Synovial membrane)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Protease K
Permeabilization
No
Specification
Synovial membrane
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 04 2017

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (brain)
Specification
brain

Abcam user community

Verified customer

Submitted Mar 03 2017

Application
Flow Cytometry
Sample
Rhesus monkey Cell (chorioretinal endothelial cell)
Permeabilization
No
Gating Strategy
size and granularity
Specification
chorioretinal endothelial cell
Preparation
Cell harvesting/tissue preparation method: cells were detached from tissue culture plates with 0.05% Trypsin-EDTA
Sample buffer: 2mM EDTA + 0.5% BSA in PBS
Fixation
Paraformaldehyde

Andrea Cabrera

Verified customer

Submitted Feb 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (Ovary)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate 10 mM, pH 6.0
Permeabilization
Yes - 0.025% Triton X-100
Specification
Ovary
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 27 2016

Application
Immunocytochemistry
Sample
Human Cultured Cells (Primary Human Internal Thoracic Artery Endothelia)
Permeabilization
Yes - Cold Methanol
Specification
Primary Human Internal Thoracic Artery Endothelia
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 25 2015

1-10 of 22 Abreviews

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