Overview

  • Product name
    Anti-CD3 antibody [CD3-12]
    See all CD3 primary antibodies
  • Description
    Rat monoclonal [CD3-12] to CD3
  • Specificity
    ab11089 recognises a highly conserved epitope of the CD3 molecule expressed by T lymphocytes.
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Horse, Chicken, Dog, Human, Pig, Rhesus monkey
    Predicted to work with: Sheep, Rabbit, Cynomolgus monkey, Woodchuck
  • Immunogen

    Synthetic peptide corresponding to CD3. Synthetic peptide derived from cytoplasmic epitope of CD3.
    Sequence:

    ERPPPVPNPDYEPC

  • Positive control
    • IHC-P:FFPE human tonsil normal.
  • General notes
    This product should be stored undiluted. Storage in frost free freezers is not recommended. Should this product contain a precipitate we recommend microcentrifugation before use.

Properties

Applications

Our Abpromise guarantee covers the use of ab11089 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.

Membrane permeabilization is recommended for this application.

ab18407 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

1mM EDTA pH8.0 is recommended for this purpose.

WB Use at an assay dependent concentration. Predicted molecular weight: 23.1 kDa.

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 [CD3-12] images

  • All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1 µg/ml

    Lane 1 : THP1 whole cell lysate (negative control)
    Lane 2 : Raji whole cell lysate (negative 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-Rat at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 23.1 kDa
    Observed band size : 23 kDa (why is the actual band size different from the predicted?)

    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 ab11089 overnight at 4°C. Antibody binding was detected using Goat anti-Rat secondary at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • IHC image of CD3 staining in a formalin fixed, paraffin embedded normal mouse lymph node tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6). The section was incubated with ab11089 at 1/250 dilution for 15 mins at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • IHC image of CD3 staining in a formalin fixed, paraffin embedded normal human tonsil tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6). The section was incubated with ab11089 at 1/250 dilution for 15 mins at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD3 with ab11089. Heat mediated antigen retrieval was performed using EDTA buffer pH 9.0.

  • Staining of human peripheral blood lymphocytes with Rat anti Human CD3 (ab11089) by Flow cytometry (FACS).

  • ab11089 staining CD3 in mouse lymph node tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 15% serum for 60 minutes at 20°C. Antigen retrieval was by heat mediation in NaCitrate (pH 6). Samples were incubated with primary antibody (1/250 in TBS) for 18 hours at 20°C. An Alexa Flour®conjugated goat anti-rat IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • Anti-CD3 antibody [CD3-12] (ab11089) + Jurkat whole cell lysate

    Secondary
    HRP-conjugated goat anti-rat IgG

    Predicted band size : 23.1 kDa

References for Anti-CD3 antibody [CD3-12] (ab11089)

This product has been referenced in:
  • Marquardt N  et al. Fetal CD103+ IL-17-Producing Group 3 Innate Lymphoid Cells Represent the Dominant Lymphocyte Subset in Human Amniotic Fluid. J Immunol 197:3069-3075 (2016). Read more (PubMed: 27591320) »
  • Burudi EM  et al. Regulation of indoleamine 2,3-dioxygenase expression in simian immunodeficiency virus-infected monkey brains. J Virol 76:12233-41 (2002). Read more (PubMed: 12414962) »

See all 3 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Thymus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate pH6.0
Permeabilization
No
Specification
Thymus
Blocking step
DakoCytomation X0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
Formaldehyde
Username

Hongwei Shao

Verified customer

Submitted Aug 28 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cynomolgus monkey Tissue sections (Lymph Node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris base/EDTA
Permeabilization
Yes - Tween 20
Specification
Lymph Node
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 24°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 14 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 50mM Borate Buffer with 1mM NaCl and 1mM EDTA - pH 8.0
Permeabilization
No
Specification
Spleen
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 23°C
Fixative
Formaldehyde
Username

Matthew Brown

Verified customer

Submitted Mar 13 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Peripheral blood mononuclear cells (PBMCs))
Permeabilization
Yes - 0.5% TX100
Specification
Peripheral blood mononuclear cells (PBMCs)
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Nov 04 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (lymph node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: NaCitrate pH 6
Permeabilization
No
Specification
lymph node
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 23 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (lymph node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: NaCitrate pH 6
Specification
lymph node
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 16 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Diva Decloaker
Sample
Human Tissue sections (Intestine)
Specification
Intestine
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 23 2014

Sorry for my delayed response to your message - I have been away from the office for a few days. We will send you a replacement vial of ab11089 on order 52778. You should already have received the order confirmation e-mail - please check to see if the ...

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I'm sorry to hear that this antibody is not working in IHC. The datasheet advises that the antibody should cross react with mouse and that it is suitable for use in IHC on PES after heat mediated antigen retrieval. Your protocol looks fine - would you ...

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Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet, these are updated as soona s any new information is brought to our attention. As far as we are aware, cross reactivity with cat has no...

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

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