Anti-CD31 antibody [MEC 7.46] (ab7388)

Reviews (6)Q&A (9)References (63)

Overview

  • Product name
    Anti-CD31 antibody [MEC 7.46]
    See all CD31 primary antibodies
  • Description
    Rat monoclonal [MEC 7.46] to CD31
  • Host species
    Rat
  • Specificity
    The antibody reacts with the murine form of the Platelet-Endothelial Cell Adhesion Molecule. The reactivity of the antibody is restricted to the isoform of the molecule that is selectively expressed by endothelial cells. The antibody precipitates a 130 kDa molecule present on the membrane of endothelial cell presents on all mouse blood vessels both in normal and inflamed or tumor tissues. The antigen is predominantly present at the lateral borders of endothelial cells as described for human PECAM-1.
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IP, IHC-Frmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Mouse t.end.1 cells (polyoma middle T (PmT)-transformed EC)

Applications

Our Abpromise guarantee covers the use of ab7388 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
Flow Cyt Use at an assay dependent concentration.

No fixation or permeabilization needed. Epitope is sensitive to trypsin!
ab18407 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration. Please see reviews for fixative recommendations. Certain users find acetone fixation necessary while others find 4% PFA necessary. In addition, found to work at 1/1000 dilution.
IP Use at an assay dependent concentration.
IHC-Fr 1/10.

Acetone (-20°C) fixation for 10 minutes at room temperature is recommended.
  • Application notes
    Is unsuitable for IHC-P.

    • 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 (Frozen sections) - Anti-CD31 antibody [MEC 7.46] (ab7388)
        Immunohistochemistry (Frozen sections) - Anti-CD31 antibody [MEC 7.46] (ab7388)This image is courtesy of an anonymous Abreview

        Immunohistochemical analysis of frozen mouse skin tissue, staining CD31 with ab7388.
        Tissue was fixed with acetone and blocked with 2% serum for 30 minutes at 20°C. Samples were incubated with primary antibody (1/300 in 1% BSA) for 4 hours at 20°C. An Alexa Fluor® 488-conjugated goat anti-rat monoclonal IgG (1/500) was used as the secondary antibody.

        See Abreview

      • Immunohistochemistry (Frozen sections) - Anti-CD31 antibody [MEC 7.46] (ab7388)
        Immunohistochemistry (Frozen sections) - Anti-CD31 antibody [MEC 7.46] (ab7388)This image is courtesy of an anonymous abreview.

        Immunohistochemistry (Frozen sections) analysis of mouse embryo transverse tissue section labeling CD31 with ab7388 at 1/1000 dilution. Tissue sections were fixed with paraformaldehyde and permeabilized with PBST (PBS / 0.5% v/v Triton X-100). Tissue was blocked for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in 1% BSA + 5% donkey serum + PBST) for 12 hours at 4°C. A polyclonal goat anti-rat Alex Fluor® was used as the secondary antibody at 1/1000 dilution. This is a transverse section of E10.5 mouse embryo. CD31 expresses in endothelium cells which locates in the surface of dorsal aorta and small vessels.

        See Abreview

      • Flow Cytometry - Anti-CD31 antibody [MEC 7.46] (ab7388)
        Flow Cytometry - Anti-CD31 antibody [MEC 7.46] (ab7388)

        Flow Cytometry analysis of bEND3 (mouse brain endothelial cell line) cells labeling CD31 with ab7388 at 10 µg/ml concentration. 106 cells were used per sample. No fixation or permeabilization needed. A R-PE labelled goat anti-rat IgG (H+L) was used as the secondary antibody.

      References

      This product has been referenced in:
      • Smolarczyk R  et al. Combination of anti-vascular agent - DMXAA and HIF-1a inhibitor - digoxin inhibits the growth of melanoma tumors. Sci Rep 8:7355 (2018). IHC-Fr ; Mouse . Read more (PubMed: 29743548) »
      • Nie X  et al. mTOR acts as a pivotal signaling hub for neural crest cells during craniofacial development. PLoS Genet 14:e1007491 (2018). Read more (PubMed: 29975682) »
      See all 63 Publications for this product

      Publishing research using ab7388? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

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      1-10 of 15 Abreviews or Q&A

      Immunohistochemistry (Frozen sections) abreview for Anti-CD31 antibody [MEC 7.46]

       Excellent
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Frozen sections)
      Sample
      Mouse Tissue sections (mouse tumor tissue)
      Permeabilization
      Yes - PBST(0.5% v/v triton X-100, sigma)
      Specification
      mouse tumor tissue
      Blocking step
      Serum 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 Dec 21 2016

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 antibody [MEC 7.46]

       Good
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Mouse Tissue sections (Adipose tissue)
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: 10mM citric acid, 55'C, 20min
      Permeabilization
      Yes - PBS/ 0.1% v/v Triton X-100 (Sigma)
      Specification
      Adipose tissue
      Blocking step
      BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
      Fixative
      Paraformaldehyde
      Read More

      Mr. Bongju Park

      Verified customer

      Submitted Dec 01 2015

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD31 antibody [MEC 7.46]

       Poor
      Abreviews
      Abcam has not validated the combination of species/application used in this Abreview.
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Mouse Tissue sections (mouse skin)
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: Dako target retrieval solution
      Permeabilization
      No
      Specification
      mouse skin
      Blocking step
      BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 24°C
      Fixative
      Paraformaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted May 19 2015

      Immunohistochemistry (Frozen sections) abreview for Anti-CD31 [MEC 7.46] antibody

       Excellent
      Abreviews
      Abcam guarantees this product to work in the species/application used in this Abreview.
      abreview image
      Application
      Immunohistochemistry (Frozen sections)
      Sample
      Mouse Tissue sections (E10.5 mouse embryos transverse section)
      Specification
      E10.5 mouse embryos transverse section
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - PBST (PBS / 0.5% v/v Triton X-100 (Sigma))
      Blocking step
      NOVOCASTRA protein block as blocking agent as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
      Read More

      Abcam user community

      Verified customer

      Submitted May 13 2013

      Immunohistochemistry (Frozen sections) abreview for Anti-CD31 [MEC 7.46] antibody

       Excellent
      Abreviews
      Abcam guarantees this product to work in the species/application used in this Abreview.
      abreview image
      Application
      Immunohistochemistry (Frozen sections)
      Sample
      Mouse Tissue sections (mouse skin)
      Specification
      mouse skin
      Fixative
      Acetone
      Permeabilization
      No
      Blocking step
      Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C
      Read More

      Abcam user community

      Verified customer

      Submitted Jan 09 2013

      Question

      Inquiry: Dear abcam-team I would like to draw your attention to one particular review submitted on 24 February 2011 in connection with the CD31-antibody ab7388. As the reviewer remained anonymous I put my question to you instead: The host of ab7388 is rat but the anonymous reviewer has used the DAKO Envision mouse polymer detection system and reported no staining. I have searched all DAKO Envision products and found that they react either with mouse and/ or with rabbit primary antibodies. This could explain those negative staining results. I am interested in purchasing ab7388 but am hesitating because of this review. Is there any chance that you could find out exactly what secondary antibody this anonymous reviewer has in fact used to detect ab7388 in his mouse tissue? I am looking forward to your reply. best regards

      Read More
      Answer

      I now have some further information to share in regards to the anti-CD31 antibody [MEC 7.46] (ab7388).

      I have been in contact with the reviewer and they have confirmed that they did indeed use an anti-mouse HRP DAKO kit to detect this rat antibody. This is likely to have yielded very poor results as the cross reactivity with rat is likely to have been poor and as thetissue being stained was from mouse the background was likely very high. Thank you for noticing this error and bringing it to our attention. I have now removed this review from the website to avoid further confusion.

      If I can help in any other way please do let me know. I wish you all the best with your research.

      Read More

      Question

      In the title of the attchament I said I used Cell cultures from bEnd3 cells. Yes, I though that the problem could be the secondary antibody but I bought it to abcam at the same time (the problem is the same) and to clarify this issue I need another primary of the same specie (rat) because I don't have it to test.   Thanks in advance,  

      Read More
      Answer

      Thank you for your response and for clarifying the cell types used in the experiments.    I understand now that bEnd3 cells (immortalized mouse brain endothelial cell line) were stained for CD31. The protocol looks absolutely fine and it you have an alternative antibody which worked fine with these cells. I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.   I could raise either a credit note for the cost of  one vial of ab7388 and ab7388 since you have alternative antibodies; or send you two replacement vials (one of each)  free of charge. Please could you let me know how you wish to proceed with your enquiry? I look forward to hearing from you soon.

      Read More

      Question

      I send in attach the protocol I used (that unswer to all questions of the inquiry). This protocol is optimized for a lot of antigens, namely CD31....   Fixative for 10 min at RT: PFA 4% Acetone PFA:Acetone (1:1) Wash 3x in PBS Permeabilize with Triton X-100 0,2%, 10 min, RT; Wash 3x in PBS Blockinh with BSA/PBS 1%, 40-60 min, RT; Wash in PBS Incubate 1st antibody, ON, 4ºC (rat CD31 abcam, ab7388) 1:50 1:100 1:500 1:1000 Wash in PBS Incubate with 2nd antibody in the dark, for 2h, RT (anti-rat abcam, ab6565) 1:1000 1:500 Wash in the dark Coverslip with Prolong Gold + DAPI. Fixative for 10 min at RT: PFA 4% Acetone PFA:Acetone (1:1) Wash 3x in PBS Permeabilize with Triton X-100 0,2%, 10 min, RT; Wash 3x in PBS Blockinh with BSA/PBS 1%, 40-60 min, RT; Wash in PBS Incubate 1st antibody, ON, 4ºC (rat CD31 abcam, ab7388) 1:50 1:100 1:500 1:1000 Wash in PBS Incubate with 2nd antibody in the dark, for 2h, RT (anti-rat abcam, ab6565) 1:1000 1:500 Wash in the dark Coverslip with Prolong Gold + DAPI.  

      Read More
      Answer

      Thank you for your response. Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem. 1)  Sample: Could you specify the samples used for the staining i.e. species, cells (cultured) or tissues? 2) Detection system: Have you used the secondary antibody successfully with another primary antibody? Do you think that the problem may come from the secondary antibody? Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

      Read More

      Immunocytochemistry/ Immunofluorescence abreview for Anti-CD31 [MEC 7.46] antibody

       Average
      Abreviews
      Abcam guarantees this product to work in the species/application used in this Abreview.
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Mouse Cell (lung tissue)
      Specification
      lung tissue
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - MeOH
      Blocking step
      Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
      Read More

      Abcam user community

      Verified customer

      Submitted Sep 10 2007

      Question

      I send you a description of my protocol. Order details: Ab 7388 Batch nr.209535 Stored at -20 deegres aliquoted. Description of the problem: No signal Paraffin-inbeded tissues. Fixed in paraformaldehyd. Heated with target ret. Buffer pH 9.9 or citric acid buffer in a micro-wave-owen. Primary ab dilution between 1/10- 1/200 Secodary ab Vectors biotinylerad rat ab. IHC in Tech-Mate- mascine ( Dakopats) including blocking, incubation with pr.ab, incation with sec.ab including envision, DAB and hematoxilin. I´ve have also tested different time for the incubation with the primary ab. Best regards

      Read More
      Answer

      Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. This antibody has not been tested using immunohistochemistry using paraffin sections. Indeed we have received various enquiries associated with its absence of reactivity against this type of fixed tissue suggesting that the antibody is not capable of binding to its antigen under these conditions. However, I would like to offer you some suggestions. This antibody has been shown to recognise mouse CD31 on frozen sections. In your enquiry you do not make reference to the type of tissue that you are using. I would like to recommend that you change the fixation of your tissue to use 5% methanol or acetone. You also do not mention the duration that you incubate for. I would like to know how long you incubate the antibody for. I would like to recommend an overnight incubation at 4oC in a dilution range of 1/100 to 1/10. I would finally like to recommend that you employ a positive control of mouse heart/lung (endothelial tissues) to help the optimisation of the antibody. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

      Read More

      1-10 of 15 Abreviews or Q&A

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