Overview

  • Product name
  • Description
    Rabbit polyclonal to CD3
  • Tested applications
    Suitable for: IP, WBmore details
    Unsuitable for: IHC
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human CD3.

    (Peptide available as ab16206.)

  • Positive control
    • This antibody gave a positive signal in Jurkat whole cells and thymus tissue from Mouse and Rat.

Properties

Applications

Our Abpromise guarantee covers the use of ab16044 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
IP Use a concentration of 5 µg/ml.
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
  • Application notes
    Is unsuitable for IHC.
  • Target

    • Function
      The CD3 complex mediates signal transduction.
    • Involvement in disease
      Defects in CD3D are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)/B(+)/NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.
    • Sequence similarities
      Contains 1 ITAM domain.
    • Cellular localization
      Membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • 4930549J05Rik antibody
      • A430104F18Rik antibody
      • AW552088 antibody
      • Cd247 antibody
      • CD247 antigen antibody
      • CD247 antigen, zeta subunit antibody
      • CD247 molecule antibody
      • CD3 antibody
      • CD3 antigen, delta subunit antibody
      • CD3 delta antibody
      • CD3 epsilon antibody
      • CD3 eta antibody
      • CD3 gamma antibody
      • CD3 molecule delta polypeptide antibody
      • CD3 molecule, epsilon polypeptide antibody
      • CD3 molecule, gamma polypeptide antibody
      • CD3 zeta antibody
      • CD3-DELTA antibody
      • CD3d antibody
      • CD3D antigen delta polypeptide antibody
      • CD3d antigen, delta polypeptide (TiT3 complex) antibody
      • CD3d molecule delta antibody
      • CD3d molecule delta CD3 TCR complex antibody
      • CD3d molecule, delta (CD3-TCR complex) antibody
      • CD3D_HUMAN antibody
      • CD3E antibody
      • CD3e antigen antibody
      • CD3E antigen epsilon polypeptide antibody
      • CD3e antigen, epsilon polypeptide (TiT3 complex) antibody
      • CD3e molecule epsilon antibody
      • CD3e molecule epsilon CD3 TCR complex antibody
      • CD3e molecule, epsilon (CD3-TCR complex) antibody
      • CD3epsilon antibody
      • CD3G antibody
      • CD3g antigen antibody
      • CD3G antigen gamma polypeptide antibody
      • CD3g antigen, gamma polypeptide (TiT3 complex) antibody
      • CD3g molecule gamma antibody
      • CD3g molecule gamma CD3 TCR complex antibody
      • CD3g molecule, gamma (CD3-TCR complex) antibody
      • CD3H antibody
      • CD3Q antibody
      • CD3Z antibody
      • CD3zeta antibody
      • Ctg3 antibody
      • FLJ17620 antibody
      • FLJ17664 antibody
      • FLJ18683 antibody
      • FLJ79544 antibody
      • FLJ94613 antibody
      • IMD19 antibody
      • Leu-4 antibody
      • MGC138597 antibody
      • OKT3, delta chain antibody
      • OTTHUMP00000032544 antibody
      • T cell receptor antibody
      • T cell receptor T3 delta chain antibody
      • T cell receptor T3 gamma chain antibody
      • T cell receptor T3 zeta chain antibody
      • T cell receptor zeta chain antibody
      • T cell surface glycoprotein CD3 antibody
      • T cell surface glycoprotein CD3 delta chain antibody
      • T cell surface glycoprotein CD3 epsilon chain antibody
      • T cell surface glycoprotein CD3 gamma chain antibody
      • T cell surface glycoprotein CD3 zeta chain antibody
      • T-cell antigen receptor complex, delta subunit of T3 antibody
      • T-cell antigen receptor complex, epsilon subunit of T3 antibody
      • T-cell antigen receptor complex, gamma subunit of T3 antibody
      • T-cell antigen receptor complex, zeta subunit of CD3 antibody
      • T-cell receptor T3 delta chain antibody
      • T-cell receptor T3 gamma chain antibody
      • T-cell surface antigen T3/Leu-4 epsilon chain antibody
      • T-cell surface glycoprotein CD3 delta chain antibody
      • T-cell surface glycoprotein CD3 gamma chain antibody
      • T3 antibody
      • T3d antibody
      • T3e antibody
      • T3g antibody
      • T3z antibody
      • TCRE antibody
      • TCRk antibody
      • Tcrz antibody
      • TCRzeta antibody
      see all

    Anti-CD3 antibody images

    • All lanes : Anti-CD3 antibody (ab16044) at 1 µg/ml

      Lane 1 : THP1 whole cell lysate (-ve control)
      Lane 2 : Raji whole cell lysate (-ve control)
      Lane 3 : Jurkat whole cell lysate
      Lane 4 : Human Thymus tissue lysate
      Lane 5 : Mouse Thymus tissue lysate
      Lane 6 : Rat Thymus tissue lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size : 23 kDa
      Observed band size : 23 kDa
      Additional bands at : 17 kDa. We are unsure as to the identity of these extra bands.

      Lanes 1 - 6: Merged signal (red and green). Green – ab16044 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.

       

      This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab16044 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

    • All lanes : Anti-CD3 antibody (ab16044) at 1 µg/ml

      Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
      Lane 2 : Thymus (Mouse) Tissue Lysate at 20 µg
      Lane 3 : Thymus (Rat) Tissue Lysate at 20 µg

      Secondary
      Anti-Rabbit IgG VHH Single Domain Antibody (HRP) (ab191866) at 1/50000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 23 kDa
      Observed band size : 23 kDa
      Additional bands at : 18 kDa,48 kDa,62 kDa. We are unsure as to the identity of these extra bands.

      Exposure time : 8 minutes

      This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab16044 overnight at 4°C. Antibody binding was detected using an anti-rabbit IgG VHH single domain antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    • CD3 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to CD3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
      The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab16044.
      Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
      Band: 23kDa; CD3

    References for Anti-CD3 antibody (ab16044)

    This product has been referenced in:
    • Daniels BP  et al. Regional astrocyte IFN signaling restricts pathogenesis during neurotropic viral infection. J Clin Invest 127:843-856 (2017). Read more (PubMed: 28134626) »
    • Zhang S  et al. Epigallocatechin-3-gallate (EGCG) inhibits imiquimod-induced psoriasis-like inflammation of BALB/c mice. BMC Complement Altern Med 16:334 (2016). IHC-P ; Mouse . Read more (PubMed: 27581210) »

    See all 7 Publications for this product

    Product Wall

    Thank you for your email.

    We have 4 rabbit antibodies that work in rat in IHC-P. I have listed them below:

    ab16044 Anti-CD3 antibody
    http://www.abcam.com/ab16044

    ab16669 Anti-CD3 antibody [SP7]
    http://www.abca...

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (Spleen)
    Permeabilization
    Yes - 0.3% TritonX in 0.1% PBS
    Specification
    Spleen
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Paraformaldehyde
    Username

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Jun 08 2011

    Abreviews
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Flow Cytometry
    Sample
    Human Cell (KM-H2)
    Specification
    KM-H2
    Preparation
    Cell harvesting/tissue preparation method: Asynchronous KM-H2 were harvested and labeled by indirect immunofluorescence
    Sample buffer: PBS + 5% FBS
    Permeabilization
    No
    Gating Strategy
    Forward/Side scatter were used to eliminate cellular debris. The accompanying marker was applied such that only 2% of the IgG control was positive.
    Username

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Dec 16 2008

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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