Overview

  • Product name

    Anti-CD226 antibody [EPR20710]
    See all CD226 primary antibodies
  • Description

    Rabbit monoclonal [EPR20710] to CD226
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse CD226 aa 1-250. The exact sequence is proprietary.
    Database link: Q8K4F0

  • Positive control

    • WB: Mouse spleen lysate; EL4.IL-2 and YAC-1 whole cell lysates. IHC-P: Mouse spleen and lung tissues. ICC/IF: EL4.IL-2 cells. IP: Mouse spleen lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab212077 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 1/1000. Detects a band of approximately 65 kDa (predicted molecular weight: 38 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/30.

Target

  • Function

    Receptor involved in intercellular adhesion, lymphocyte signaling, cytotoxicity and lymphokine secretion mediated by cytotoxic T-lymphocyte (CTL) and NK cell.
  • Tissue specificity

    Expressed by peripheral blood T-lymphocytes.
  • Sequence similarities

    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • adhesion glycoprotein antibody
    • CD226 antibody
    • CD226 antigen antibody
    • CD226 molecule antibody
    • CD226_HUMAN antibody
    • DNAM 1 antibody
    • DNAM-1 antibody
    • DNAM1 antibody
    • DNAX accessory molecule 1 antibody
    • platelet and T cell activation antigen 1 antibody
    • PTA1 antibody
    • T lineage specific activation antigen 1 antigen antibody
    • TLiSA1 antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized EL4.IL-2 (mouse lymphoma T lymphocyte cell line) cells labeling CD226 with ab212077 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)(ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on EL4.IL-2 cells. 

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • All lanes : Anti-CD226 antibody [EPR20710] (ab212077) at 1/1000 dilution

    Lane 1 : Mouse spleen lysate
    Lane 2 : EL4.IL-2 (mouse lymphoma T lymphocyte cell line) whole cell lysate
    Lane 3 : YAC-1 (mouse Moloney murine leukemia virus induced lymphoma lymphoblast cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG H&L (HRP))

    Developed using the ECL technique.

    Predicted band size: 38 kDa
    Observed band size: 65 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lanes 1-2: 1 minute; Lane 3: 3 minutes.

    The different molecular weights (~65kDa) observed in different tissues or cell lines may be due to different level of CD226 glycosylation (PMID: 8673704).

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD226 with ab212077 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Cytoplasmic staining on megakaryocytes and platelets of mouse spleen is observed (PMID: 15693793). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), Ready to use

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling CD226 with ab212077 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Positive staining on platelets of mouse lung is observed (PMID: 15693793). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), Ready to use.

  • CD226 was immunoprecipitated from 0.35 mg of mouse spleen lysate with ab212077 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab212077 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

    Lane 1: Mouse spleen lysate 10 μg (Input).

    Lane 2: ab212077 IP in mouse spleen lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab212077 in mouse spleen lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds.

References

ab212077 has not yet been referenced specifically in any publications.

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