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Immunohistochemistry-Paraffin: CD3 Antibody (SP7) [NB600-1441] - FFPE human tonsil stained with CD3 antibody.
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 ...read more
Genetic Strategies: 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 ...read more
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).
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.
Isotype
IgG
Clonality
Monoclonal
Host
Rabbit
Gene
CD3E
Purity
Tissue culture supernatant
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.
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).
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.
Use in Mouse reported in scientific literature (PMID:35111697). Canine reactivity per customer review. Use in Porcine reported in scientific literature (PMID:33839961)
Packaging, Storage & Formulations
Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
Tissue culture supernatant
Preservative
0.05% Sodium Azide
Purity
Tissue culture supernatant
Alternate Names for CD3 Antibody (SP7)
CD_antigen: CD3e
CD3 antigen, delta subunit
CD3
CD3d antigen
CD3d antigen, delta polypeptide (TiT3 complex)
CD3d molecule, delta (CD3-TCR complex)
CD3-DELTA
CD3e antigen
CD3e antigen, epsilon polypeptide (TiT3 complex)
CD3e molecule, epsilon (CD3-TCR complex)
CD3e
CD3-epsilon
CD3g antigen
CD3g antigen, gamma polypeptide (TiT3 complex)
CD3g molecule, epsilon (CD3-TCR complex)
CD3g molecule, gamma (CD3-TCR complex)
CD3G
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
Background
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
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
Limitations
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.
Yotis DY DM Differential Effects of KIM-1 in Subcutaneous and Orthotopic Renca Models of Kidney Cancer Thesis Jan 1 2021
Maisonneuve C, Tsang D, Foerster E Et Al. Nod1 Promotes Colorectal Carcinogenesis by Regulating the Immunosuppressive Functions of Tumor-Infiltrating Myeloid Cells SSRN Journal Jun 26 2020
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
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.
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Diseases for CD3 Antibody (NB600-1441)
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