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RecombinantRabMAb

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

Submit a review Submit a question References (1)
Immunocytochemistry/ Immunofluorescence - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
  • Immunoprecipitation - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
  • Western blot - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
  • Western blot - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
  • Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22667-12] to CD3 epsilon
  • Suitable for: WB, IHC-P, ICC/IF, IP
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

Carrier Free

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. This information is proprietary to Abcam and/or its suppliers.

  • 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

    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.

Properties

Applications

The Abpromise guarantee

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.
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).

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

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (1)

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

    ab231775 has been referenced in 1 publication.

    • Yi M  et al. Combine and conquer: manganese synergizing anti-TGF-ß/PD-L1 bispecific antibody YM101 to overcome immunotherapy resistance in non-inflamed cancers. J Hematol Oncol 14:146 (2021). PubMed: 34526097

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