Recombinant
RabMAb

Recombinant Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)

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Overview

  • Product name

    Anti-CD3 epsilon antibody [EPR22667-12]
    See all CD3 epsilon primary antibodies

  • Description

    Rabbit monoclonal [EPR22667-12] to CD3 epsilon

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Recombinant fragment within Mouse CD3 epsilon aa 1-100. The exact sequence is proprietary.
    Database link: P22646

  • Positive control

    • WB: EL4.IL2 whole cell lysate; mouse thymus lysate. IP: EL4.IL2 whole cell lysate. IHC-P: Mouse spleen and lung tissue. ICC/IF: EL4.IL2 cells.

  • 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 ab231775 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 18, 22 kDa (predicted molecular weight: 21 kDa).
IHC-P 1/5000. 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.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      The CD3 complex mediates signal transduction.

    • Sequence similarities

      Contains 1 Ig-like (immunoglobulin-like) domain.
      Contains 1 ITAM domain.

    • Cellular localization

      Membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • CD3 epsilon antibody
        • CD3e antibody
        • CD3e antigen antibody
        • CD3e antigen epsilon polypeptide (TiT3 complex) antibody
        • CD3E antigen epsilon polypeptide antibody
        • CD3E antigen, epsilon subunit antibody
        • CD3e molecule epsilon antibody
        • CD3e molecule, epsilon (CD3 TCR complex) antibody
        • CD3e molecule, epsilon (CD3-TCR complex) antibody
        • CD3E_HUMAN antibody
        • IMD18 antibody
        • T cell antigen receptor complex epsilon subunit of T3 antibody
        • T cell surface antigen T3/Leu 4 epsilon chain antibody
        • T cell surface glycoprotein CD3 epsilon chain antibody
        • T-cell surface antigen T3/Leu-4 epsilon chain antibody
        • T-cell surface glycoprotein CD3 epsilon chain antibody
        • T3E antibody
        • TCRE antibody
        see all

      Images

      • Immunocytochemistry/ Immunofluorescence - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
        Immunocytochemistry/ Immunofluorescence - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)

        Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized EL4.IL2 (mouse lymphoma T lymphocyte cell line) cells labeling CD3 epsilon with ab231775 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 and cytoplasmic staining in EL4.IL2 cell line is observed. 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, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)

        Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3 epsilon with ab231775 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the mouse spleen (PMID: 18648532) is observed. 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.

        Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      • Immunoprecipitation - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
        Immunoprecipitation - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)

        CD3 epsilon was immunoprecipitated from 0.35 mg of EL4.IL2 (mouse lymphoma T lymphocyte cell line) whole cell lysate with ab231775 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab231775 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as the secondary antibody at 1/5000 dilution.

        Lane 1: EL4.IL2 whole cell lysate 10 μg (Input).
        Lane 2: ab231775 IP in EL4.IL2 whole cell lysate.
        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab231775 in EL4.IL2 whole cell lysate.

        Blocking and dilution buffer and concentration: 5% NFDM/TBST.
        Exposure time: 30 seconds.

      • Western blot - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
        Western blot - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
        Anti-CD3 epsilon antibody [EPR22667-12] (ab231775) at 1/1000 dilution + EL4.IL2 (mouse lymphoma T lymphocyte cell line) whole cell lysate at 20 µg

        Secondary
        Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

        Predicted band size: 21 kDa
        Observed band size: 22 kDa why is the actual band size different from the predicted?


        Exposure time: 158 seconds


        Blocking and dilution buffer: 5% NFDM/TBST.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)

        Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling CD3 epsilon with ab231775 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the T lymphocytes on mouse lung (PMID: 22888136) is observed. 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.

        Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      • Western blot - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
        Western blot - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
        Anti-CD3 epsilon antibody [EPR22667-12] (ab231775) at 1/1000 dilution + Mouse thymus lysate at 10 µg

        Secondary
        Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 21 kDa
        Observed band size: 18,22 kDa why is the actual band size different from the predicted?


        Exposure time: 15 seconds


        Blocking and dilution buffer: 5% NFDM/TBST.

        The expression profile observed is consistent with what has been described in the literature (PMID: 1387664).

      References

      ab231775 has not yet been referenced specifically in any publications.

      Publishing research using ab231775? Please let us know so that we can cite the reference in this datasheet.

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