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    cd31-antibody-ab32457.pdf

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Anti-CD31 antibody (ab32457)

  • Datasheet
  • SDS
Reviews (11)Q&A (3)References (63)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (ab32457)
  • Immunocytochemistry - Anti-CD31 antibody (ab32457)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (ab32457)
  • Western blot - Anti-CD31 antibody (ab32457)
  • Immunocytochemistry - Anti-CD31 antibody (ab32457)
  • Flow Cytometry - Anti-CD31 antibody (ab32457)

Key features and details

  • Rabbit polyclonal to CD31
  • Suitable for: ICC, Flow Cyt, IHC-P, WB
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-CD31 antibody [EPR3094] (ab76533)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-CD31 antibody
    See all CD31 primary antibodies
  • Description

    Rabbit polyclonal to CD31
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC, Flow Cyt, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human CD31 aa 600-700 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab32456)

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Blood
    • Platelets
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Cell Adhesion Molecules
    • Endothelial
    • Cardiovascular
    • Angiogenesis
    • Angiogenic Factors
    • Stem Cells
    • Mesenchymal Stem Cells
    • Surface Molecules
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Cell adhesion
    • Other
    • Cancer
    • Tumor immunology
    • CD markers
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Cell adhesion molecules
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Monocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Neutrophil Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Thrombocytic Lineage
    • Cardiovascular
    • Cardiovascular Markers
    • Cell Markers
    • Endothelial Cells
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human CD31 protein (ab155607)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32457 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 (3)
Use a concentration of 5 µg/ml.

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

Flow Cyt (1)
Use 0.1-1µg for 106 cells.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P (3)
1/2000.

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

WB (2)
Use a concentration of 1 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 130 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).

Notes
ICC
Use a concentration of 5 µg/ml.

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

Flow Cyt
Use 0.1-1µg for 106 cells.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P
1/2000.

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

WB
Use a concentration of 1 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 130 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).

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.
  • 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

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (ab32457)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (ab32457)
      IHC image of ab32457 staining in human tonsil formalin fixed paraffin embedded tissue section, 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 20 mins. The section was then incubated with ab32457, 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
    • Immunocytochemistry - Anti-CD31 antibody (ab32457)
      Immunocytochemistry - Anti-CD31 antibody (ab32457)

      ab32457 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 (ab32457 at 1µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (ab32457)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (ab32457)This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.

      ab32457 staining CD31 in human tonsil tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in Citrate buffer pH 6.0 and blocking (5 minutes/peroxidase block then 10 minutes/protein block) for 15 minutes at 20°C. The primary antibody was diluted, 1/2000 and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.

      See Abreview

    • Western blot - Anti-CD31 antibody (ab32457)
      Western blot - Anti-CD31 antibody (ab32457)
      All lanes : Anti-CD31 antibody (ab32457) at 1 µg/ml

      Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 2 : HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate
      Lane 3 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Performed under reducing conditions.

      Predicted band size: 130 kDa
      Observed band size: 130 kDa
      Additional bands at: 26 kDa, 55 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 30 seconds
    • Immunocytochemistry - Anti-CD31 antibody (ab32457)
      Immunocytochemistry - Anti-CD31 antibody (ab32457)

      ICC image of ab32457 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32457, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Flow Cytometry - Anti-CD31 antibody (ab32457)
      Flow Cytometry - Anti-CD31 antibody (ab32457)

      Overlay histogram showing Jurkat cells stained with ab32457 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32457, 0.1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (polyclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (63)

    Publishing research using ab32457? Please let us know so that we can cite the reference in this datasheet.

    ab32457 has been referenced in 63 publications.

    • Offeddu GS  et al. The cancer glycocalyx mediates intravascular adhesion and extravasation during metastatic dissemination. Commun Biol 4:255 (2021). PubMed: 33637851
    • Ng WL  et al. Fabrication and Characterization of 3D Bioprinted Triple-layered Human Alveolar Lung Models. Int J Bioprint 7:332 (2021). PubMed: 33997432
    • Han X  et al. Analysis of glucose metabolism by 18F-FDG-PET imaging and glucose transporter expression in a mouse model of intracerebral hemorrhage. Sci Rep 11:10885 (2021). PubMed: 34035344
    • Hedegaard CL  et al. Peptide-protein coassembling matrices as a biomimetic 3D model of ovarian cancer. Sci Adv 6:N/A (2020). PubMed: 33008910
    • Zhang R  et al. LncRNA TUG1 regulates autophagy-mediated endothelial-mesenchymal transition of liver sinusoidal endothelial cells by sponging miR-142-3p. Am J Transl Res 12:758-772 (2020). PubMed: 32269710
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 14 Abreviews or Q&A

    IHC - Wholemount abreview for Anti-CD31 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Human Tissue (HUVEC spheroid)
    Specification
    HUVEC spheroid
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 15 2020

    Western blot abreview for Anti-CD31 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human umbilical vein endothelial cell)
    Gel Running Conditions
    Reduced Denaturing (12 %)
    Loading amount
    20 µg
    Specification
    Human umbilical vein endothelial cell
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 15 2020

    Immunocytochemistry abreview for Anti-CD31 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cell (Liver Sinusoidal Endothelial cells)
    Permeabilization
    No
    Specification
    Liver Sinusoidal Endothelial cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 15 2020

    Western blot abreview for Anti-CD31 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Pig Tissue lysate - whole (skin)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Treatment
    burn wound with stem cell treatment
    Specification
    skin
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 20 2015

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Vector Antigen Unmasking Solution
    Permeabilization
    No
    Specification
    Tumor
    Blocking step
    Peroxidase as blocking agent for 5 minute(s) · Concentration: 3% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 26 2013

    Immunocytochemistry abreview for Anti-CD31 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cell (Human brain microvascular endothelial cell)
    Permeabilization
    Yes - 1% Triton X-100
    Specification
    Human brain microvascular endothelial cell
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 20 2012

    Immunocytochemistry abreview for Anti-CD31 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cell (normal prostate)
    Permeabilization
    Yes - 0.5mg/ml saponin in 1% BSA
    Specification
    normal prostate
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 27°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Satyendra Tripathi

    Verified customer

    Submitted Aug 10 2012

    Question

    I wonder what is the difference between PECAM primary antibody
    ab24590 and ab32457, why one of them detected PECAM protein molecular weight on 83kD, the other one detected130kD.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 29 2013

    Answer



    It is an interesting situation and I'm glad you mentioned it. The theoretical mass of CD31 is about 86kDa. However CD31 can be extensively modified post-translationally. These modifications include phosphorylation, palmitoylation and glycosylation. The degree of modification may cause CD31 to run slower than expected, which is why there may be a smeary band around 130kDa.

    I agree it is confusing when an 'expected weight' is listed as both 86kDa and 130kDa. However I believe the post-translational modifications are the reason for listing 130kDa as an expected molecular weight.

    Read More

    Abcam Scientific Support

    Answered on Apr 29 2013

    Question

    Has the reactivity of ab32457 against mouse samples been tested? in this case the customer needs to know for sure it doesn't react with Mouse derived samples.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 09 2013

    Answer

    >I can confirm that ab32457 has not yet been tested in mouse for cross-reactivity.

    Read More

    Abcam Scientific Support

    Answered on Jan 09 2013

    Question

    I would like to know why your cd31 rabbit antibody (ab32457 ) stains epithelial cells (HeLa). I thought CD31 was specific for endothelial cells.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 16 2012

    Answer

    Looking through the literature, it seems that CD31 is primarily expressed on platelets, leukocytes, and endothelial cells. It is also expressed on some tumor cell lines-

    http://www.ncbi.nlm.nih.gov/pubmed/16518857?dopt=Abstract

    It has also been shown to be an adhesion molecule expressed on HeLa cells-

    http://www.ncbi.nlm.nih.gov/pubmed/9269301?dopt=Abstract

    Due to this expression, HeLa cells and lysates are commonly used as a positive control for CD31 antibodies.

    Read More

    Abcam Scientific Support

    Answered on Nov 16 2012

    1-10 of 14 Abreviews or Q&A

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