Overview

  • Product name
    Anti-CD3 antibody [SP7]
    See all CD3 primary antibodies
  • Description
    Rabbit monoclonal [SP7] to CD3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, IHC-P, WB, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Sheep, Rabbit, Horse, Chicken, Cow, Cat, Dog, Baboon, Woodchuck
  • Immunogen

    Synthetic peptide corresponding to CD3.

  • Epitope
    aa 156-168 of epsilon chain of Human CD3 protein (intracytoplasmic).
  • Positive control
    • WB: Jurkat whole cell lysate. Human, mouse and rat thymus tissue lysate. IHC-P: Pig and rat spleen tissue. Human tonsil tissue. Mouse epidydimal fat pad tissue. Rat infarcted heart tissue. Flow Cytometry: Human peripheral blood lymphocytes. Jurkat cells. IHC-Fr: Mouse brain tissue.
  • General notes

    This antibody is suitable for staining normal and neoplastic T cells in formalin-fixed, paraffin-embedded tissues.

Properties

Applications

Our Abpromise guarantee covers the use of ab16669 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 1/1000.

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

We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody.

IHC-Fr Use at an assay dependent concentration. PubMed: 18658050
IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.

WB 1/200. Predicted molecular weight: 23 kDa.
IHC-FoFr Use at an assay dependent concentration.

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

Images

  • Immunohistochemical analysis of Formaldehyde fixed, paraffin-embedded pig spleen tissue sections labelling CD3 with ab16669 at a dilution of 1/100. Biotin conjugated Goat Anti-Rabbit IgG was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid.

    See Abreview

  • Immunohistochemical analysis of Formaldehyde fixed, frozen rat spleen tissue sections labelling CD3 with ab16669 at a dilution of 1/500. Biotin conjugated Goat Anti-Rabbit IgG at 1/300 dilution was used as the secondary antibody. 

    Cryostat sections (10 microns thick) of fresh frozen spleen were dried overnight. They were fixed in 10% Formalin in PBS pH7 for 15 minutes.
    After secondary antibody incubation ( Goat anti-rabbit biotin) , streptavidin-Alexa 594 was applied.

    See Abreview

  • All lanes : Anti-CD3 antibody [SP7] (ab16669) at 1/25 dilution

    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
    All lanes : 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



    Lanes 1 - 6: Merged signal (red and green). Green – ab16669 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 ab16669 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • Immunohistochemical (Formalin / PFA fixed paraffin-embedded sections) staining of of Tonsil tissue labeling CD3 (SP7) with ab16669 at 1/150 dilution.

  • Immunohistochemical analysis of Formaldehyde fixed, paraffin-embedded rat spleen tissue sections labelling CD3 with ab16669 at a dilution of 1/100. Biotin conjugated Goat Anti-Rabbit IgG at 1/300 dilution was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid.

    See Abreview

  • Ab16669 staining CD3 in Mouse Epidydimal fat pad tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections). Tissue sections were fixed with formaldehyde, blocked with 5% serum for 4 hours at 25°C and permeabilized with Triton X-100. Samples were incubated with primary antibody (1/100 in PBST with BSA and goat serum) for 4°C at 12 hours. An Alexa Fluor® 568 goat anti-rabbit IgG (H + L) cross adsorbed was used as the secondary antibody.

    See Abreview

  • Human peripheral blood lymphocytes stained with ab16669 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab16669, 1/1000 dilution) for 30 min at 4°C. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.

  • Flow cytometric analysis of rabbit anti-CD3 (SP7) antibody ab16669 (1/100) in Jurkats cells (green) compare to negative control of rabbit IgG (blue).

  • Human tonsil stained with ab16669.
  • ab16669 staining rat infarcted heart tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    Myocardial infarction was produced in a rat model following the ligation of the left anterior descending (LAD) coronary artery. Tissue was harvested 6 w following infarct, fixed with Histochoice for 72 hr, paraffin sectioned and the slide was then baked prior to CD3 staining. ab16669 at 1/200 was incubated overnight at 4°C. The image was taken with a confocal laser scanning microscope and shows cells giving strong inmmunofluorescence staining for CD3 antigen (green), indicating presence of cells of T-lymphocytes origin in the infarct zone of the heart tissue, counterstained nuclei with DAPI (blue). Note, CD3 tended to be present in nests of 2-5 cells that were non-uniformly distributed in the infarct zone. In addition, the image shows that the CD3 localization is predominantly membrane based and to a certain extent intracytoplasmic.

    See Abreview

  • Anti-CD3 antibody [SP7] (ab16669) at 1/25 dilution + Jurkat cell lysate

    Predicted band size: 23 kDa
    Observed band size: 19 kDa (why is the actual band size different from the predicted?)

  • ab16669 staining CD3 in murine brain tissue by Immunohistochemistry (Frozen sections).
    Tissue was fixed in acetone, blocked using 5% serum for 30 minutes at 25°C and then incubated with ab16669 at a 1/200 dilution for 18 hours at 4°C. The secondary used was an undiluted HRP conjugated goat anti-rabbit polyclonal.

    See Abreview

  • Immunohistochemical analysis of Human tonsil tissue, staining CD3 (green) with ab16669.

    Antigen retrieval was performed by heat mediation in citrate buffer (pH 6) and blocked with 5% goat serum and 5% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/100) overnight at 4°C. A Cy3®-conjugated anti-rabbit IgG was used as the secondary antibody.

References

This product has been referenced in:
  • Luo N  et al. DNA methyltransferase inhibition upregulates MHC-I to potentiate cytotoxic T lymphocyte responses in breast cancer. Nat Commun 9:248 (2018). Read more (PubMed: 29339738) »
  • Höing B  et al. Stromal versus tumoral inflammation differentially contribute to metastasis and poor survival in laryngeal squamous cell carcinoma. Oncotarget 9:8415-8426 (2018). IHC-P . Read more (PubMed: 29492204) »

See all 130 Publications for this product

Customer reviews and Q&As

can confirm that ab16669 CD3 antibody [SP7] is sold as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody conce...

Read More

Thank you for contacting us.

I am sorry to confirm that because we carry over 90,000 products, it is not feasible for us to stock small sample sizes of our products to send on request.

We are happy to reassure our customers that a...

Read More

I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application us...

Read More

Thank you for your enquiry.

I can confirm that the Anti-CD3 antibody [SP7] (ab16669) antibody isprovided as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will ...

Read More

Thank you for contacting us.

We currently do not offer products which meet each of the requirements that you have set. I assembled for you well characterized products which meet most of these requirements but each has some deficiency. Pric...

Read More

Thank you very much for your phone call.

The best antibodies we can recommend are as follows. Please note that the recommendations are based on immunogen sequence only we currently do not hold any rabbit specific data.

Anti CD3 : ab...

Read More

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

Thank you for your enquiry. According to the on-line datasheet, this antibody is recommended to be stored at -20°C and it should be viable for up to 5 years. However, it is important to emphasize that our Abpromise guarantees that the pr...

Read More

Thank you for contacting us. The relative specificity of each antibody is difficult to assess without a side-by-side comparison. The data for the SP7 clone and the positive reviews we have received from customers are all encouraging. St...

Read More

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

Sign up