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CD3 Antibody (SP7)

Novus Biologicals | Catalog # NB600-1441

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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Porcine, Canine

Cited:

Human, Mouse, Porcine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Rabbit IgG Clone # SP7
View all CD3 Primary Antibodies »

Product Specifications

Immunogen

This CD3 antibody was developed against a synthetic peptide: KAKAKPVTRGAGA, corresponding to amino acids 156-168 of Human CD3 epsilon chain.

Epitope

aa 156-168 of epsilon chain of human CD3 protein (intracytoplasmic).

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:35111697). Canine reactivity per customer review. Use in Porcine reported in scientific literature (PMID:33839961)

Localization

Type I membrane protein.

Specificity

The CD3 antigen is present on early thymocytes and mature T cells and is generally regarded as a pan-T cell marker. This antibody will help detect CD3 expression in normal and neoplastic tissues. This antibody reacts with the intracytoplasmic portion of the CD3 antigen expressed by T cells. It stains human T cells in both the cortex and medulla of the thymus and in peripheral lymphoid tissues. This antibody is suitable for staining normal and neoplastic T cells in formalin-fixed, paraffin-embedded tissues.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

23.1 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for CD3 Antibody (SP7)

Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441]

Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441]

Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441] - FFPE human tonsil stained with CD3 antibody.
Immunohistochemistry-Frozen: CD3 Antibody (SP7) [NB600-1441]

Immunohistochemistry-Frozen: CD3 Antibody (SP7) [NB600-1441]

Immunohistochemistry-Frozen: CD3 Antibody (SP7) [NB600-1441] - T-cells and microglia in mouse spinal cord with acute EAE. Mouse was perfused with 4% paraformaldehyde. Spinal cord was post-fixed overnight, followed by cryoprotection with 30% sucrose for 24h. Spinal cord sections were treated with antigen-retrieval buffer (pH 6.0) at 60C for 10 min, then cooled down to room temperature (this step should not be omitted). Anti-CD3 antibody was diluted at 1:200. Incubation: room temperature overnight. IHC-Fr image submitted by a verified customer review.
Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441]

Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441]

Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441] - Mouse lung tissue stained against C3aR normal conditions (left) and knockdown conditions (right). IHC-P image submitted by a verified customer review.

Applications for CD3 Antibody (SP7)

Application
Recommended Usage

Immunohistochemistry

1:25-1:50

Immunohistochemistry-Frozen

1:10 - 1:500

Immunohistochemistry-Paraffin

1:25-1:50
Application Notes
IHC-P: recommended pretreatment of citrate buffer, pH 6.0. Recommended incubation time of 30-60 min at RT. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 29037255). Use in FLOW reported in scientific literature (PMID: 31079916).

Reviewed Applications

Read 5 reviews rated 3.6 using NB600-1441 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Tissue culture supernatant

Formulation

Tissue culture supernatant

Preservative

0.05% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: CD3

CD3 (cluster of differentiation marker 3) is a multi-subunit transmembrane protein that is expressed on the surface of T-cells and forms a complex with the T-cell receptor (TCR) (1-3). CD3 consists of four distinct membrane protein isoforms: CD3-delta (delta), CD3-epsilon (epsilon), CD3-gamma, and CD3-zeta (1-3). The CD3 subunits organize into a complex containing three sets of dimers: CD3-epsilondelta, CD3-epsilongamma, CD3-zetazeta. The CD3 complex binds to the TCR heterodimer (alphabeta or gammadelta) to form the transmembrane TCR-CD3 complex (2-3). Structurally, the CD3 protein chains have an extracellular region, a transmembrane domain, and a cytoplasmic trail region (2-3). The CD3-epsilondelta and CD3-epsilongamma heterodimers also contain an extracellular immunoglobulin (Ig)-like domain, classifying them as part of the immunoglobulin superfamily (2-3). The cytoplasmic tail of each CD3 chain also contains one (delta, epsilon, gamma) or three (zeta) immunoreceptor tyrosine-based activation motifs (ITAMs), for a total of 10 ITAMs in the whole CD3 complex (2-3). Following, TCR binding to peptide major histocompatibility complex (p-MHC), the CD3 ITAMs are phosphorylated by the Src kinase Lck and are important for recruiting ZAP70 and initiating TCR signaling cascade activation (2). While similar in structure, the CD3 chains vary in length and molecular weight (4). The CD3-delta is 171 amino acids (aa) in length with a theoretical molecular weight of 18.9 kDa (4, 5). The CD3-epsilon is 204 aa long and has a theoretical molecular weight of 23 kDa (4, 5). CD3-gamma is 182 aa long with a theoretical molecular weight of 20.4 kDa (4, 5). Finally, CD3-zeta is 164 aa in length with a theoretical molecular weight of 18.6 kDa (4, 5).

CD3 proteins are expressed on the surface of thymocytes during thymocyte development, proliferation, and maturation to T-cells (4, 6, 7). During T-cell development CD4-CD8- double negative (DN) cells differentiate to CD4+CD8+ double positive (DP) cells before progressing to single positive (SP) CD4+ helper T-cells or CD8+ cytotoxic T-cells (4, 6, 7). As CD3 plays an important role in thymocyte development, it is understandable that CD3 defects and mutations in CD3 protein chains cause severe combined immunodeficiencies (SCIDs) (8). Additionally, a subset of CD3+ T-cells that co-express CD20 are described in a variety of diseases including rheumatoid arthritis, multiple sclerosis, CD20+ T-cell leukemia/lymphoma, and HIV (9). Clinical trials and animal models have shown that anti-CD3 monoclonal antibodies are a promising treatment modality for inflammatory disorders and autoimmune diseases, such as type I diabetes (10).

References

1. Chetty, R., & Gatter, K. (1994). CD3: structure, function, and role of immunostaining in clinical practice. The Journal of pathology. https://doi.org/10.1002/path.1711730404

2. Mariuzza, R. A., Agnihotri, P., & Orban, J. (2020). The structural basis of T-cell receptor (TCR) activation: An enduring enigma. The Journal of biological chemistry. https://doi.org/10.1074/jbc.REV119.009411

3. Kuhns, M. S., Davis, M. M., & Garcia, K. C. (2006). Deconstructing the form and function of the TCR/CD3 complex. Immunity. https://doi.org/10.1016/j.immuni.2006.01.006

4. Clevers, H., Alarcon, B., Wileman, T., & Terhorst, C. (1988). The T cell receptor/CD3 complex: a dynamic protein ensemble. Annual review of immunology. https://doi.org/10.1146/annurev.iy.06.040188.003213

5. Uniprot: CD3-delta (P04234), CD3-epsilon (P07766), CD3-gamma (P09693), CD3-zeta (P20963)

6. D'Acquisto, F., & Crompton, T. (2011). CD3+CD4-CD8- (double negative) T cells: saviours or villains of the immune response?. Biochemical pharmacology. https://doi.org/10.1016/j.bcp.2011.05.019

7. Dave V. P. (2009). Hierarchical role of CD3 chains in thymocyte development. Immunological reviews. https://doi.org/10.1111/j.1600-065X.2009.00835.x

8. Fischer, A., de Saint Basile, G., & Le Deist, F. (2005). CD3 deficiencies. Current opinion in allergy and clinical immunology. https://doi.org/10.1097/01.all.0000191886.12645.79

9. Chen, Q., Yuan, S., Sun, H., & Peng, L. (2019). CD3+CD20+ T cells and their roles in human diseases. Human immunology. https://doi.org/10.1016/j.humimm.2019.01.001

10. Kuhn, C., & Weiner, H. L. (2016). Therapeutic anti-CD3 monoclonal antibodies: from bench to bedside. Immunotherapy. https://doi.org/10.2217/imt-2016-0049

Alternate Names

CD_antigen: CD3e, CD3 antigen, delta subunit, CD3d antigen, CD3d antigen, delta polypeptide (TiT3 complex), CD3d molecule, delta (CD3-TCR complex), CD3-DELTA, CD3e, CD3e antigen, CD3e antigen, epsilon polypeptide (TiT3 complex), CD3e molecule, epsilon (CD3-TCR complex), CD3-epsilon, CD3G, CD3g antigen, CD3g antigen, gamma polypeptide (TiT3 complex), CD3g molecule, epsilon (CD3-TCR complex), CD3g molecule, gamma (CD3-TCR complex), CD3-GAMMA, FLJ17620, FLJ17664, FLJ18683, FLJ79544, FLJ94613, IMD18, MGC138597, T3DOKT3, delta chain, T3E, T-cell antigen receptor complex, epsilon subunit of T3, T-cell receptor T3 delta chain, T-cell surface antigen T3/Leu-4 epsilon chain, T-cell surface glycoprotein CD3 delta chain, T-cell surface glycoprotein CD3 epsilon chain, TCRE

Entrez Gene IDs

916 (Human); 917 (Human); 915 (Human)

Gene Symbol

CD3E

Additional CD3 Products

Product Documents for CD3 Antibody (SP7)

Certificate of Analysis

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Product Specific Notices for CD3 Antibody (SP7)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for CD3 Antibody (SP7)

Customer Reviews for CD3 Antibody (SP7) (5)

3.6 out of 5
5 Customer Ratings
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4 Stars
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1 Stars
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Customer Images

Showing  1 - 5 of 5 reviews Showing All
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  • CD3 Antibody (SP7)
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Spleen
    Species: Mouse
    Verified Customer | Posted 03/19/2024
    CD3 immunoreactivity in sections of mouse FFPE spleen obtained with varying dilutions of NB600-1441. Primary was left on tissue sections for 30m at room temperature.
    Target Retrieval Solution was used for heat induced epitope retrieval using a vegetable steamer.
    CD3 Antibody (SP7) NB600-1441
    Bio-Techne Response
    Thank you for reviewing our product. We are sorry to hear that this product did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.
  • CD3 Antibody (SP7)
    Name: Kevin Thorburn
    Application: Immunohistochemistry-Paraffin
    Sample Tested: FFPE
    Species: Dog
    Verified Customer | Posted 04/06/2023
    NB600-1441 at 1 to 50 dilution for 1h at room temperature produces weak labeling in dog spleen. Recommend clone CD3-12 instead for detection of dog CD3 in FFPE sections.
    Tested a variety of concentrations from 1 to 25 to 1 to 800 with an incubation time of 1h at room temperature. NB600-1441 did not produce any signal in any of the 4 sections of dog spleen that were used for testing. The sections were known to contain CD3, as clone CD3-12 produces excellent labeling.
    CD3 Antibody (SP7) NB600-1441
    Bio-Techne Response
    Thank you for reviewing our product. We are sorry to hear that this product did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.
  • CD3 Antibody (SP7)
    Name: Tyler Crowe
    Application: Immunohistochemistry-Frozen
    Sample Tested: Frozen human skin sections
    Species: Human
    Verified Customer | Posted 08/20/2018
    CD3 (red, 1:100) stained in frozen human skin section. Counterstained with DAPI
    Human skin section fixed with PFA, and permeablized with 0.5% Triton. Sections were blocked with 5% BSA, before anti-CD3 ab was applied at 1:100 for 1hr at 37C. anti-rb-A568 secondary was used, and counterstained with DAPI
    CD3 Antibody (SP7) NB600-1441
  • CD3 Antibody (SP7)
    Name: Xuenong Bo
    Application: Immunohistochemistry-Frozen
    Sample Tested: mouse spinal cord cryo-sections
    Species: Mouse
    Verified Customer | Posted 02/21/2018
    T-cells and microglia in mouse spinal cord with acute EAE
    Mouse was perfused with 4% paraformaldehyde. Spinal cord was post-fixed overnight, followed by cryoprotection with 30% sucrose for 24h. Spinal cord sections were treated with antigen-retrieval buffer (pH 6.0) at 60C for 10 min, then cooled down to room temperature (this step should not be omitted). Anti-CD3 antibody was diluted at 1:200. Incubation: room temperature overnight.
    CD3 Antibody (SP7) NB600-1441
  • CD3 Antibody (SP7)
    Name: Hans Snyder
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Human Tonsil & Canine lymph paraffin slides
    Species: Human and Canine
    Verified Customer | Posted 08/25/2017
    Human tonsil
    Works great in both Human and canine tissues. Sodium citrate pH 6.0 Concentration 1:100
    CD3 Antibody (SP7) NB600-1441

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