RecombinantRabMAb

Recombinant Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

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Flow Cytometry - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
  • Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
  • Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
  • Immunoprecipitation - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
  • Dot Blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
  • Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP776(2)Y] to CD3 zeta (phospho Y83)
  • Suitable for: Flow Cyt, ICC/IF, Dot blot, WB, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free FITC

Overview

  • Product name

    Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y]
    See all CD3 zeta primary antibodies

  • Description

    Rabbit monoclonal [EP776(2)Y] to CD3 zeta (phospho Y83)

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human CD3 zeta aa 50-150 (phospho Y83). The exact sequence is proprietary.
    Database link: P20963

  • Positive control

    • WB: Jurkat whole cell lysate (ab7899). IP: Jurkat. ICC: Jurkat cells. Flow Cyt: Jurkat 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.

    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. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EP776(2)Y

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab68236 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.
ICC/IF
1/100 - 1/250.
Dot blot
Use at an assay dependent concentration.
WB
1/5000 - 1/10000. Detects a band of approximately 18-22 kDa (predicted molecular weight: 18 kDa).
IP
1/50.
Notes
Flow Cyt
Use at an assay dependent concentration.
ICC/IF
1/100 - 1/250.
Dot blot
Use at an assay dependent concentration.
WB
1/5000 - 1/10000. Detects a band of approximately 18-22 kDa (predicted molecular weight: 18 kDa).
IP
1/50.
Application notes
Is unsuitable for IHC-P.

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.
  • Information by UniProt

  • Database links

    • 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

    • Flow Cytometry - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      Flow Cytometry - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

      Flow Cytometry analysis of Jurkat (human acute T cell leukemia) treated (Red)/untreated (Green) with 1mM pervanadate for 4 hours with purified ab68236 at 1/250 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

    • Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      All lanes : Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236) at 1/2000 dilution

      Lane 1 : Untreated Jurkat cells whole cell lysates
      Lane 2 : Jurkat cells were treated with 50mM Pervanadate for 5 minutes whole cell lysates
      Lane 3 : Jurkat cells were treated with 50mM Pervanadate for 5 minutes whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 18 kDa
      Observed band size: 18 kDa


      Exposure time: 3 minutes


      Blocking buffer 5% NFDM/TBST

      Diluting buffer 5% NFDM/TBST

    • Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      All lanes : Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236) at 1/10000 dilution

      Lane 1 : Jurkat cell lysate, untreated.
      Lane 2 : Jurkat cell lysate, treated with pervanadate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

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

    • Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

      Immunocytochemistry/Immunofluorescence analysis of Jurkat cells (untreated, Per treated and Per+LP treated) labelling CD3 zeta (phospho Y83) with ab68236 (left) and CD3 zeta with ab40804 (right) both at a dilution of 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      The image shows increased cytoplamic staining after Pervanadate (1 mM, 30 min) treatment on Jurkat cells. The LP treatment decreased the cytoplasmic staining caused by Pervanadate.

      ab40804 was used as a Pan control for ab68236. The results showed cytoplamic staining on untreated, pervanadate (1 mM, 30 min) treated and Per+LP treated Jurkat cells.

    • Immunoprecipitation - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      Immunoprecipitation - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

      CD3 zeta was immunoprecipitated from 0.35 mg Jurkat (Human T cell leukemia T lymphocyte) treated with pervandate (50mM 5min) whole cell lysate 10 µg with ab68236 at 1/30 dilution (2µg) . VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Jurkat (Human T cell leukemia T lymphocyte) treated with pervandate (50mM 5min) whole cell lysate 10 µg

      Lane 2: ab68236 IP in Jurkat treated with pervandate (50mM 5min) whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab68236 in Jurkat treated with pervandate (50mM 5min) whole cell lysate

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

    • Dot Blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      Dot Blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

      Dot blot analysis of CD3 zeta (pY83) phospho peptide (lane 1) and CD3 zeta non-phospho peptide (lane 2) labelling CD3 zeta (phospho Y83) with ab68236 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
      Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)

    References (14)

    Publishing research using ab68236? Please let us know so that we can cite the reference in this datasheet.

    ab68236 has been referenced in 14 publications.

    • Eleftheriadis T  et al. A Role for Human Renal Tubular Epithelial Cells in Direct Allo-Recognition by CD4+ T-Cells and the Effect of Ischemia-Reperfusion. Int J Mol Sci 22:N/A (2021). PubMed: 33572206
    • 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
    • Ramello MC  et al. An immunoproteomic approach to characterize the CAR interactome and signalosome. Sci Signal 12:N/A (2019). PubMed: 30755478
    • Razvag Y  et al. T Cell Activation through Isolated Tight Contacts. Cell Rep 29:3506-3521.e6 (2019). PubMed: 31825832
    • Quintarelli C  et al. Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma. Oncoimmunology 7:e1433518 (2018). PubMed: 29872565
    View all Publications for this product

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