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RecombinantRabMAb

Recombinant Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)

Submit a review Submit a question References (6)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Western blot - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Immunocytochemistry - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Western blot - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • OI-RD Scanning - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
  • Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)

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 - BSA and Azide free
  • Suitable for: WB, ICC, IHC-P
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Biotin Unconjugated

Overview

  • Product name

    Anti-CD31 antibody [EPR3094] - BSA and Azide free
    See all CD31 primary antibodies

  • Description

    Rabbit monoclonal [EPR3094] to CD31 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC, 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: Jurkat cells

  • General notes

    ab207090 is the carrier-free version of ab76533.

    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.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab207090 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
WB
Use at an assay dependent concentration. 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).
ICC
Use at an assay dependent concentration.

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
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

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
WB
Use at an assay dependent concentration. 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).
ICC
Use at an assay dependent concentration.

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
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

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.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

    • Western blot - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Western blot - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      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



      Blocking and diluting buffer: 5% NFDM /TBST 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

    • Immunocytochemistry - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Immunocytochemistry - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Immunocytochemistry analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling CD31 with ab76533 at 1/500 (4.9 μg/mL). ab150077, AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/mL) was used as the secondary antibody. Cells were fixed with 100% Methanol. DAPI (blue) was used as nuclear counterstain.

      Confocal image showing positive staining on Jurkat cells.
       
      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

       

    • Western blot - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Western blot - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      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



      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)

      Clone EPR3094 (ab207090) has been successfully conjugated by Abcam. This image was generated using Anti-CD31 antibody [EPR3094] (Alexa Fluor® 647). Please refer to ab218582 for protocol details.

      IHC image of CD31 staining in a section of formalin-fixed paraffin-embedded normal human kidney*.

      The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Biocare Medical NxGen pressure cooker using retrieval settings of 110oC for 8 minutes. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature.  The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab218582 at 1/100 dilution (shown in red) and counterstained using ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

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

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

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)

      Immunohistochemical analysis of paraffin embedded human kidney tissue using ab76533 at a 1/250 dilution. Note positive staining of endothelial cells.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

      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] - BSA and Azide free (ab207090)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)

      Immunohistochemical analysis of paraffin embedded human muscle tissue using ab76533 at a 1/250 dilution. Note positive staining of endothelial cells.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

      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] - BSA and Azide free (ab207090)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)This image is 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

    • OI-RD Scanning - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      OI-RD Scanning - Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)

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

      Click here to learn more about KD

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76533).

    • Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)
      Anti-CD31 antibody [EPR3094] - BSA and Azide free (ab207090)

    References (6)

    Publishing research using ab207090? Please let us know so that we can cite the reference in this datasheet.

    ab207090 has been referenced in 6 publications.

    • Xue Y  et al. Tumor-infiltrating M2 macrophages driven by specific genomic alterations are associated with prognosis in bladder cancer. Oncol Rep 42:581-594 (2019). PubMed: 31233191
    • Ren L  et al. Preparation of Three-Dimensional Vascularized MSC Cell Sheet Constructs for Tissue Regeneration. Biomed Res Int 2014:301279 (2014). IHC-P, ICC ; Mouse, Human . PubMed: 25110670
    • He KF  et al. CD163+ tumor-associated macrophages correlated with poor prognosis and cancer stem cells in oral squamous cell carcinoma. Biomed Res Int 2014:838632 (2014). IHC-P ; Human . PubMed: 24883329
    • Shu Q  et al. Vasostatin inhibits VEGF-induced endothelial cell proliferation, tube formation and induces cell apoptosis under oxygen deprivation. Int J Mol Sci 15:6019-30 (2014). ICC ; Human . PubMed: 24722573
    • Hofmann NA  et al. Oxygen sensing mesenchymal progenitors promote neo-vasculogenesis in a humanized mouse model in vivo. PLoS One 7:e44468 (2012). IHC-P ; Human . PubMed: 22970226
    • Balcells M  et al. Smooth muscle cells orchestrate the endothelial cell response to flow and injury. Circulation 121:2192-9 (2010). PubMed: 20458015

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