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    cd3-zeta-phospho-y142-antibody-ep2652y-ab68235.pdf

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Immunology Adaptive Immunity T Cells CD
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RecombinantRabMAb

Recombinant Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)

  • Datasheet
  • SDS
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Western blot - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
  • Immunocytochemistry - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
  • Immunoprecipitation - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
  • Flow Cytometry (Intracellular) - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
  • Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP265(2)Y] to CD3 zeta (phospho Y142)
  • Suitable for: Flow Cyt (Intra), ICC, WB, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y]
    See all CD3 zeta primary antibodies
  • Description

    Rabbit monoclonal [EP265(2)Y] to CD3 zeta (phospho Y142)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC, WB, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • Jurkat whole cell lysate (ab7899)
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP265(2)Y
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD

Associated products

  • Alternative Versions

    • Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] - BSA and Azide free (ab245765)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat membrane extract lysate (ab30130)
  • Recombinant Protein

    • Recombinant Human CD3 zeta protein (ab139254)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab68235 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 (Intra)
1/120.
ICC
1/250 - 1/500.
WB
1/500 - 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 19 kDa).
IP
1/20 - 1/50.
Notes
Flow Cyt (Intra)
1/120.
ICC
1/250 - 1/500.
WB
1/500 - 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 19 kDa).
IP
1/20 - 1/50.

Target

  • Function

    Probable role in assembly and expression of the TCR complex as well as signal transduction upon antigen triggering.
  • Involvement in disease

    Defects in CD247 are the cause of immunodeficiency due to defect in CD3-zeta (CD3ZID) [MIM:610163]. An immunological deficiency characterized by T-cells impaired immune response to alloantigens, tetanus toxoid and mitogens.
  • Sequence similarities

    Belongs to the CD3Z/FCER1G family.
    Contains 3 ITAM domains.
  • Domain

    The ITAM domains mediate interaction with SHB.
  • Post-translational
    modifications

    Phosphorylated on Tyr residues after T-cell receptor triggering.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession P20963 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 919 Human
    • Omim: 186780 Human
    • SwissProt: P20963 Human
    • Unigene: 156445 Human
    • Alternative names

      • 4930549J05Rik antibody
      • A430104F18Rik antibody
      • AW552088 antibody
      • CD247 antibody
      • CD247 antigen antibody
      • Cd247 molecule antibody
      • Cd3 antibody
      • CD3 antigen, zeta polypeptide, isoform CRA_b antibody
      • CD3 antigen, zeta subunit antibody
      • CD3-eta antibody
      • CD3H antibody
      • CD3Q antibody
      • Cd3z antibody
      • CD3Z antigen zeta polypeptide (TiT3 complex) antibody
      • CD3Z_HUMAN antibody
      • CD3zeta antibody
      • CD3zeta chain antibody
      • MGC140430 antibody
      • T cell receptor T3 zeta chain antibody
      • T cell surface glycoprotein CD3 zeta chain antibody
      • T-cell antigen receptor complex, zeta subunit of CD3 antibody
      • T-cell receptor T3 zeta chain antibody
      • T-cell surface glycoprotein CD3 zeta chain antibody
      • T3z antibody
      • TCR zeta chain antibody
      • TCRk antibody
      • TCRZ antibody
      • TCRzeta antibody
      see all

    Images

    • Western blot - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
      Western blot - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
      All lanes : Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235) at 1/1000 dilution (Purified)

      Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
      Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) treated with 50mM pervanadate for 5 minutes whole cell lysates

      Lysates/proteins at 15 µg per lane.

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

      Predicted band size: 19 kDa
      Observed band size: 19 kDa

    • Immunocytochemistry - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
      Immunocytochemistry - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)

      Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) treated with 1mM pervanadate for 30 minutes cells labeling CD3 zeta with purified ab68235 at 1/500 dilution (0.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Immunoprecipitation - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
      Immunoprecipitation - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)

      ab68235 (purified) at 1/20 dilution (0.5ug) immunoprecipitating CD3 zeta in Jurkat treated with 50nM pervanadate for 5 minutes whole cell lysate whole cell lysate. Jurkat (Human T cell leukemia T lymphocyte) treated with 50nM pervanadate for 5 minutes whole cell lysate 10ug
      Lane 2 (+): ab68235 & Jurkat treated with 50nM pervanadate for 5 minutes whole cell lysate whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68235 in Jurkat treated with 50nM pervanadate for 5 minutes whole cell lysate
      For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/2000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Flow Cytometry (Intracellular) - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
      Flow Cytometry (Intracellular) - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)

      Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) treated with 50nM pervanadate for 5 minutes cells labeling CD3 zeta with purified ab68235 at 1/120 dilution (1µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). Untreated Jurkat cells (Green). 

       

       

    • Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
      Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (8)

    Publishing research using ab68235? Please let us know so that we can cite the reference in this datasheet.

    ab68235 has been referenced in 8 publications.

    • Ma Y  et al. Clustering of the ?-Chain Can Initiate T Cell Receptor Signaling. Int J Mol Sci 21:N/A (2020). PubMed: 32429097
    • Sun C  et al. THEMIS-SHP1 Recruitment by 4-1BB Tunes LCK-Mediated Priming of Chimeric Antigen Receptor-Redirected T Cells. Cancer Cell 37:216-225.e6 (2020). PubMed: 32004441
    • Caratelli S  et al. In vitro elimination of epidermal growth factor receptor-overexpressing cancer cells by CD32A-chimeric receptor T cells in combination with cetuximab or panitumumab. Int J Cancer 146:236-247 (2020). PubMed: 31479522
    • Weber EW  et al. Pharmacologic control of CAR-T cell function using dasatinib. Blood Adv 3:711-717 (2019). PubMed: 30814055
    • Lee J  et al. Induction of Immunosuppressive CD8+CD25+FOXP3+ Regulatory T Cells by Suboptimal Stimulation with Staphylococcal Enterotoxin C1. J Immunol 200:669-680 (2018). WB ; Human . PubMed: 29237775
    • Schaer DA  et al. The CDK4/6 Inhibitor Abemaciclib Induces a T Cell Inflamed Tumor Microenvironment and Enhances the Efficacy of PD-L1 Checkpoint Blockade. Cell Rep 22:2978-2994 (2018). WB . PubMed: 29539425
    • Li L  et al. Ionic CD3-Lck interaction regulates the initiation of T-cell receptor signaling. Proc Natl Acad Sci U S A 114:E5891-E5899 (2017). PubMed: 28659468
    • Pageon SV  et al. Functional role of T-cell receptor nanoclusters in signal initiation and antigen discrimination. Proc Natl Acad Sci U S A 113:E5454-63 (2016). ICC/IF . PubMed: 27573839

    Customer reviews and Q&As

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