Recombinant

Recombinant Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (ab251594)

Overview

  • Product name

    Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free
    See all CD3 epsilon primary antibodies
  • Description

    Rabbit monoclonal [CAL54] to CD3 epsilon - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human CD3 epsilon aa 150-250. The exact sequence is proprietary.
    Database link: P07766

  • Positive control

    • IHC-P: Human tonsil, gastric carcinoma and NSCLC tissue. ICC/IF: Jurkat cells. Flow cyt: Human PBMCs. IP: Human thymus lysate; Jurkat whole cell lysate.
  • General notes

    Ab251594 is the carrier-free version of ab237707. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab251594 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab251594 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.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for WB.
  • 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

    • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD3 epsilon with ab237707 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the human tonsil is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      The section was incubated with ab237707 for 30 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

    • Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling CD3 epsilon with ab237707 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the infiltrating T lymphocytes in the human gastric carcinoma is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      The section was incubated with ab237707 for 30 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling CD3 epsilon with ab237707 at 1/55 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in Jurkat cells. 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).

      PBS 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

    • CD3 epsilon was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab237707 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237707 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000  dilution.

      Lane 1: Jurkat whole cell lysate 10 μg (Input). 
      Lane 2: ab237707 IP in Jurkat whole cell lysate. 
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237707 in Jurkat whole cell lysate.

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

    • Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween 20 permeabilized human PBMCs (peripheral blood mononuclear cells) labeling CD3 epsilon with ab237707 at 1/500 (Right compared with a Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left). Goat Anti-Rabbit IgG Fc (Alexa Fluor® 488) preadsorbed (ab150097), at 1/5000 dilution was used as the secondary antibody.

      Cells were surface stained with anti-CD3 epsilon conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min followed by intracellular staining rabbit IgG (Left) or ab237707 (Right).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

    • CD3 epsilon was immunoprecipitated from 0.35 mg of human thymus lysate with ab237707 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237707 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000  dilution.

      Lane 1: Human thymus lysate 10 μg (Input). 
      Lane 2: ab237707 IP in human thymus lysate. 
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237707 in human thymus lysate.

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

    • Formalin-fixed, paraffin-embedded human NSCLC tissue stained for CD3 epsilon using ab237707 at 0.5 µg/mL in immunohistochemcial analysis.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

    • Formalin-fixed, paraffin-embedded human tonsil tissue stained for CD3 epsilon using ab237707 at 0.5 µg/mL in immunohistochemcial analysis.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

    References

    ab251594 has not yet been referenced specifically in any publications.

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