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    cd3-epsilon-antibody-epr53612-ab133628.pdf

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Immunology Adaptive Immunity T Cells CD
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RecombinantRabMAb

Recombinant Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)

  • Datasheet
  • SDS
Submit a review Q&A (1)References (1)

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Immunocytochemistry/ Immunofluorescence - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
  • Flow Cytometry - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
  • Flow Cytometry - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
  • Western blot - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
  • Immunoprecipitation - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
  • Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5361(2)] to CD3 epsilon
  • Suitable for: ICC/IF, WB, IP, Flow Cyt
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

APC Carrier Free PE

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Protein
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Recombinant human CD3 epsilon protein (Active) (ab167755)
Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Primary
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Alexa Fluor® 555 Anti-CD3D antibody [EP4426] (ab208514)

View more associated products

Overview

  • Product name

    Anti-CD3 epsilon antibody [EPR5361(2)]
    See all CD3 epsilon primary antibodies
  • Description

    Rabbit monoclonal [EPR5361(2)] to CD3 epsilon
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IP, Flow Cytmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • Jurkat and Human thymus lysates, HuT78 cells IP: Hut-78 whole cell lysate.
  • 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.

    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 -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5361(2)
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Stem Cells
    • Hematopoietic Progenitors
    • Hematopoietic Stem Cells
    • Murine Lineage Negative
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage

Associated products

  • Alternative Versions

    • Anti-CD3 epsilon antibody [EPR5361(2)] - BSA and Azide free (ab240074)
  • Isotype control

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

    • Jurkat membrane extract lysate (ab30130)
  • Recombinant Protein

    • Recombinant human CD3 epsilon protein (Active) (ab167755)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133628 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
ICC/IF
Use at an assay dependent concentration.
WB
1/10000 - 1/50000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
IP
1/10 - 1/100.
Flow Cyt
1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Notes
ICC/IF
Use at an assay dependent concentration.
WB
1/10000 - 1/50000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
IP
1/10 - 1/100.
Flow Cyt
1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Application notes
Is unsuitable for IHC-P.

Target

  • Function

    The CD3 complex mediates signal transduction.
  • Sequence similarities

    Contains 1 Ig-like (immunoglobulin-like) domain.
    Contains 1 ITAM domain.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession P07766 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 916 Human
    • Omim: 186830 Human
    • SwissProt: P07766 Human
    • Unigene: 3003 Human
    • 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 [EPR5361(2)] (ab133628)
      Immunocytochemistry/ Immunofluorescence - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed Jurkat (human T cell leukemia T lymphocyte) cells labelling CD3 epsilon with ab133628 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Jurkat cells. Counterstained with Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1:200 dilution.
      The negative controls was Raji cell line.

    • Flow Cytometry - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
      Flow Cytometry - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)

      Flow cytometric analysis of Human peripheral blood mononuclear cells (PBMC) labelling CD3 with ab133628 at 1/100 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (left) and an unlabelled control (cells without incubation with primary antibody and secondary antibody). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

    • Flow Cytometry - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
      Flow Cytometry - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)

      Flow cytometry overlay histogram of Jurkat (Human T cell leukemia T lymphocyte) cells labelling CD3 epsilon  with ab133628 at 1/100 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

    • Western blot - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
      Western blot - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
      All lanes : Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628) at 1/1000 dilution (Anti-CD3 epsilon antibody)

      Lane 1 : Human peripheral blood mononuclear cell (PBMC) lysate with NFDM/TBST
      Lane 2 : Human thymus tissue lysate with NFDM/TBST
      Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate with NFDM/TBST

      Lysates/proteins at 20 µg per lane.

      Blocking peptides at 5 % per lane.

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

      Predicted band size: 23 kDa
      Observed band size: 23 kDa


      Exposure time: 10 seconds
    • Immunoprecipitation - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
      Immunoprecipitation - Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)

      Purified ab133628 at 1/50 dilution (2µg) immunoprecipitating CD3 epsilon in Hut-78 whole cell lysate.
      Lane 1 (input): Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate 10µg
      Lane 2 (+): ab133628 + Hut-78 whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133628 in Hut-78 whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 23 kDa

    • Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)
      Anti-CD3 epsilon antibody [EPR5361(2)] (ab133628)

    Protocols

    • Western blot protocols
    • Flow cytometry protocols
    • Immunoprecipitation protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (1)

    Publishing research using ab133628? Please let us know so that we can cite the reference in this datasheet.

    ab133628 has been referenced in 1 publication.

    • 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

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Question



    Do you know what color secondary antibody was used, and do
    you have a recommended secondary to use with this antibody? Also would
    it be possible to receive a sample size of this product and negative
    control before we go about purchasing an entire vial? The assay we
    are optimizing has very spread uses so if it works we will need lots
    of this antibody!

    Read More

    Abcam community

    Verified customer

    Asked on Feb 26 2013

    Answer



    A FITC goat anti-rabbit IgG (H+L) secondary was used to do the experiment.

    Read More

    Abcam Scientific Support

    Answered on Feb 28 2013

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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