Overview

  • Product name

    Anti-CD31 antibody [RM0032-1D12]
    See all CD31 primary antibodies
  • Description

    Rat monoclonal [RM0032-1D12] to CD31
  • Host species

    Rat
  • Tested applications

    Suitable for: IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Tissue, cells or virus corresponding to CD31. Mouse endothelial membrane protein

  • Positive control

    • IHC-P: Mouse kidney, lung, heart and liver tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab56299 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-P 1/50 - 1/400.

4% PFA ix preferred fixative over formalin. Fixation should be overnight. Carrying out Proteinase K enzymatic retrieval is essential (please see specific protocol below and Abreviews). A sensitive detection system such as HRP polymer system is recommended.

IHC-Fr Use at an assay dependent concentration. PubMed: 23269798
ICC/IF Use at an assay dependent concentration. PubMed: 24474690

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

  • Tumor vascularization and VEGF in ascites.

    (A) ID8, ID8-c-MYC, or ID8-KRAS cells (2 × 106) were intra-peritoneal injected into C57/BL6 mice. Mice were sacrificed when the body weight reached 23 g. Disseminations were obtained from mice when sacrificed. Tumor vascularization of each dissemination was assessed using CD31 immunohistochemistry (ab56299).

    Scale bars represent 200 μm and 50 μm at low and high magnification, respectively.

    Paraffin sections (4-μm-thick) of the biggest tumor sections from ID8, ID8-c-MYC, and ID8-KRAS mice were dewaxed in xylene and rehydrated through graded ethanol to water. Antigens were retrieved by boiling in 10 mM citrate buffer (pH 6.0) for 30 min. The cooled sections were incubated in a peroxidase blocking solution for 10 min to quench endogenous peroxidase. Sections were incubated in a protein blocking solution at room temperature for 10 min to block non-specific binding. Sections were then stained for Ki67 using rabbit monoclonal antibody against mouse Ki67, CD31 using a rat monoclonal antibody against mouse CD31 (ab56299, Abcam 1:100 dilution).

  • Proliferation and cell number in male wild-type (+/y) and male Mct8 knockout (−/y) placentae.

    (Panel C): Representative picture demonstrating CD31 immunoreactivity (marker of endothelial cells represented by brown staining) in the mouse placenta at E18.5. The nuclei were counterstained using hematoxylin. Endothelial cells are indicated by arrows and trophoblast cells by arrowheads.

  • ab56299 at 1/400 staining CD31 in LPS treated mouse kidney tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).

  • ab56299 at 1/400 dilution, staining CD31 in LPS treated mouse liver tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).

  • ab56299 at 1/400 staining CD31 in LPS treated mouse lung tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).

  • ab56299 at 1/400 dilution, staining CD31 in LPS treated mouse heart tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections).

References

This product has been referenced in:

  • Kim AJ  et al. Endothelial Cell Lineage Analysis Does Not Provide Evidence for EMT in Adult Valve Homeostasis and Disease. Anat Rec (Hoboken) 302:125-135 (2019). Read more (PubMed: 30306735) »
  • Cortes E  et al. Tamoxifen mechanically reprograms the tumor microenvironment via HIF-1A and reduces cancer cell survival. EMBO Rep 20:N/A (2019). Read more (PubMed: 30538116) »
See all 35 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - nuclear (liver)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jul 13 2015

Question
Answer

The antibody was raised against a mouse endothelial membrane preparation, and as it is a monoclonal antibody, it could target any region of the protein. The epitope has not been mapped, unfortunately, so we do not know whether it targets one of the termini or an internal region.

Read More

Answer

On the bottom of the online datasheet for ab56299 there is a "Recommended IHC-P protocol with ab56299" link under the "Protocols" heading. In summation:

1.Dewax and Rehydration: Place the slides in a rack and sequentially pass the rack through following solutions: Histoclear or Xylene twice, 10 minutes each; and then a graded series of 100%, 90%, 70%, and 50% ethanol, 2 minutes each.
2. Wash the slides with tap water for 5 minutes and once with DI water.
3. Antigen Retrieval: treat the slides with 20µg/ml proteinase K in PBS for 15-30 minutes at room temperature.
4. Wash the slides with tap water for 5 minutes and quench the endogenous peroxidase treated in 3% H2O2 in DI water for 30 minutes
5. Wash the slides with tap water for 5mins and soaking in PBS-Tween20 (0.1%)
6. Block the slides with blocking buffer (2.5-25% BSA in PBS-Tween20) for 10 minutes
7. Add primary antibody (Rat against mouse CD31 1:100) and incubate the antibody for 2 hours at room temperature, or for overnight at 4°C.
8. Wash the slides with rotation in PBS-Tween20 for 3 times, 5 minutes each.
9. Add anti rat secondary antibody and incubate at room temperature following manufacturer instructions.
10. Wash the slides with rotation in PBS-Tween20 for 3 times, 5 minutes each.
11. Add Polymer-HRP labeled anti rabbit (Dako: K4010) and incubate for 30 minutes at room temperature.
12. Wash the slides with rotation in PBS-Tween20 for 3 times, 5 minutes each.
13. Perform DAB Color development (reagents available in Dako K4010).. 15. Wash with tap water and count stain the nuclear with hematoxin.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Proteinase K
Permeabilization
Yes - Triton X
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Kate Schunke

Verified customer

Submitted Aug 20 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Specification
kidney
Fixative
Paraformaldehyde
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: proteinase K
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Mar 06 2012

Answer

Ab6703 should work as the secondary antibody in the amplification step.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Heart Frozen Section)
Specification
Heart Frozen Section
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 26 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Skeletal muscle)
Specification
Skeletal muscle
Fixative
10% Formalin
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Proteinase K, 10ug/ml, 15 min RT
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Mr. Josh Meisner

Verified customer

Submitted Apr 08 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Human melanoma tumor recovered from nude mice.)
Specification
Human melanoma tumor recovered from nude mice.
Fixative
Formaldehyde
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: proteinase K 10ug/ml RT 30min
Permeabilization
No
Blocking step
DakoCytomation Serum Free General Blocking Cat#X0909 as blocking agent for 30 minute(s) · Concentration: 0% · Temperature: RT°C

Hongwei Shao

Verified customer

Submitted Oct 22 2009

1-10 of 11 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up