CD3 Antibody (N26-R)

Novus Biologicals | Catalog # NBP1-79054

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # N26-R
View all CD3 Primary Antibodies »

Product Specifications

Immunogen

This CD3 antibody was developed against a peptide derived from cytoplasmic, C-terminal region of human CD3-epsilon chain. Antibody recognizes the epitope between Lys156 - Glu178.

Epitope

Lys156 - Glu178

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.

Description

This antibody is immunoaffinity purified with immunogenic peptide as a ligand.

Scientific Data Images for CD3 Antibody (N26-R)

Immunohistochemistry-Paraffin: CD3 Antibody (N26-R) [NBP1-79054]

Immunohistochemistry-Paraffin: CD3 Antibody (N26-R) [NBP1-79054]

Immunohistochemistry-Paraffin: CD3 Antibody (N26-R) [NBP1-79054] - CD3 positivity in T-lymphocytes of the normal lymph node, stained with anti-CD3 antibody. FFPE human tissue (4 um section) stained.

Applications for CD3 Antibody (N26-R)

Application
Recommended Usage

Immunohistochemistry

1:100 -1:200

Immunohistochemistry-Paraffin

1:100 - 1:200
Application Notes
Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID: 24695686).

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

20mM Tris-HCl (pH 8.0) and 20mg/ml BSA

Preservative

0.05% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

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

Stability & Storage

Store at 4C. Do not freeze.

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

Gene Symbol

CD3E

UniProt

P07766

Additional CD3 Products

Product Documents for CD3 Antibody (N26-R)

Certificate of Analysis

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Download SDS

Product Specific Notices for CD3 Antibody (N26-R)

This antibody is immunoaffinity purified with immunogenic peptide as a ligand.

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 (N26-R)

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Protocols

View specific protocols for CD3 Antibody (N26-R) (NBP1-79054):

IHC-P Protocol (NBP1-79054):
1. Deparaffinize the section in 3 changes of xylene, 5 minutes each.
2. Wash the section in 96%, 80% and 70% benzyl alcohol for 5 minutes each.
3. Rinse in distilled water.
4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
5. Wash in distilled water.
6. For antigen retrieval: immerse the slide in Tris-EDTA buffer, pH 9.0, 0.05% Tween- 20*, and incubate in
microwave (850W) for 20 minutes. (Alternatively adjust to your own protocol, keeping the required pH)
7. Remove the staining to room temperature and let the slide to cool for 15 minutes.
8. Rinse in distilled water.
9. Wash in 0.05 M Tris-HCl, pH 7.6 buffer supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes.
10. Incubate the section with primary antibody diluted in buffer A at the dilution 1:100 - 200 for 1 hour in the closed
wet chamber.
11. Wash twice 5 minutes with buffer A.
12. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard
immunohistochemistry protocol (HRP - Peroxide - DAB).
13. Wash twice 5 minutes with buffer A.
14. Apply the chromogen (DAB), 10 minutes.
15. Wash in water - 10 minutes.
16. Stain in hematoxylin for 5 minutes.
17. Wash in water - 10 minutes.
18. Dehydrate the section in 2 changes of 96% benzyl alcohol for 5 minutes each.
19. Wash the section in 2 changes of xylene for 2 minutes each.
20. Mount the slide for observation.

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FAQs

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