Overview

  • Product name
  • Description
    Rabbit polyclonal to CD31
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Synthetic peptide within Mouse CD31 aa 650 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab155816)

  • Positive control
    • This antibody gave a positive signal in both Mouse Heart and Spleen tissue lysates and TH2 whole cell lysate in Western Blot as well as Mouse Placenta formalin fixed paraffin embedded tissue section in Immunohistochemistry.

Properties

Applications

Our Abpromise guarantee covers the use of ab124432 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 a concentration of 1 µg/ml. Detects a band of approximately 120-140 kDa (predicted molecular weight: 81 kDa).

Abcam recommends using milk as the blocking agent - 3%

IHC-P Use a concentration of 1 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

  • All lanes : Anti-CD31 antibody (ab124432) at 1 µg/ml

    Lane 1 : TH2 whole cell lysate
    Lane 2 : Mouse spleen

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 81 kDa
    Observed band size : 120-140 kDa (why is the actual band size different from the predicted?)

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab124432 overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • IHC image of CD31 staining in mouse placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH6 for 20 mins. The section was then incubated with ab124432, 1µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.

  • All lanes : Anti-CD31 antibody (ab124432) at 1 µg/ml (Milk blocking - 3%)

    Lane 1 : Heart (Mouse) Tissue Lysate
    Lane 2 : Spleen (Mouse) Tissue Lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 81 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 90 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutesAbcam recommends using milk as the blocking agent - 3%. CD31 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab124432 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

This product has been referenced in:
  • Yudasaka M  et al. Near-Infrared Photoluminescent Carbon Nanotubes for Imaging of Brown Fat. Sci Rep 7:44760 (2017). Read more (PubMed: 28317858) »
  • Zhang D  et al. Engineering biomimetic periosteum with ß-TCP scaffolds to promote bone formation in calvarial defects of rats. Stem Cell Res Ther 8:134 (2017). Read more (PubMed: 28583167) »

See all 7 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (muscle)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrat-Buffer pH 6,0
Permeabilization
No
Specification
muscle
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 30 2016

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (uterus)
Specification
uterus
Username

Dr. Athilakshmi Kannan

Verified customer

Submitted Jun 22 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (lung)
Loading amount
20 µg
Specification
lung
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 12 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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