Key Product Details

Species Reactivity

Human, Mouse

Applications

CyTOF-ready, Flow (Cell Surface), Flow Cytometry, Functional, Immunoblotting, In vitro assay, In vivo assay

Label

Alexa Fluor 488 (Excitation = 488 nm, Emission = 515-545 nm)

Antibody Source

Monoclonal Mouse IgG2a Kappa Clone # OKT3

Concentration

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

View all CD3 Antibodies »

Product Specifications

Immunogen

This CD3 antibody was developed against human PBMCs.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 25485679)

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2a Kappa

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 (OKT3) [Alexa Fluor® 488]

Flow Cytometry: CD3 Antibody (OKT3) [Alexa Fluor (R) 488] [NBP2-24867AF488] - A cell surface stain was performed on hPBMCs with CD8 Alpha antibody (RPA-T8) NBP2-25195 (top image) and a matched isotype control NBP2-27287 (bottom image). Cells were incubated in an antibody dilution of 1:200 for 20 minutes at room temperature. Both antibodies were conjugated to PerCP. A co-stain was also performed using CD3 antibody NBP2-24867AF488.
Flow (Cell Surface): CD3 Antibody (OKT3) [Alexa Fluor (R) 488] [NBP2-24867AF488] - Analysis of CD3 expression in hPBMc's gated to lymphocyte population at 1 x 10^6 cells/ml. Cells were stained with NBP2-24867AF488 (orange) conjugate at a 1:500 dilution. Results are shown with mIgG2 (AF488 conjugate) isotype control (blue)

Applications for CD3 Antibody (OKT3) [Alexa Fluor® 488]

Application
Recommended Usage

CyTOF-ready

Optimal dilutions of this antibody should be experimentally determined.

Flow (Cell Surface)

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Functional

Optimal dilutions of this antibody should be experimentally determined.

Immunoblotting

Optimal dilutions of this antibody should be experimentally determined.

In vitro assay

Optimal dilutions of this antibody should be experimentally determined.

In vivo assay

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Optimal dilution of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

50mM Sodium Borate

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 in the dark.

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, Muromonab

Gene Symbol

CD3E

Additional CD3 Products

Product Documents for CD3 Antibody (OKT3) [Alexa Fluor® 488]

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Download SDS

Product Specific Notices for CD3 Antibody (OKT3) [Alexa Fluor® 488]

Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

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.

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