CD3 antibody

Catalog No. ABIN114137

Product Details anti-CD3 Antibody

Target: CD3

Reactivity: Mouse

Host: Hamster

Clonality: Monoclonal

Conjugate: This CD3 antibody is un-conjugated

Application: Flow Cytometry (FACS), Immunoprecipitation (IP), Immunohistochemistry (Frozen Sections) (IHC (fro)), Functional Studies (Func), Western Blotting (WB)

Specificity: This anti-Mouse CD3e monoclonal antibody is specific for a 25 kDa protein component (e-T3) of the antigen specific T cell receptor on all Mouse strains tested. The epsilon-T3 protein has been shown to be non-covalently associated on the cell surface ab heterodimer of the CD3 associated complex. This monoclonal antibody reacts with all mature T cells and can both activate and inhibit T cell function (1). This fact identifies epsilon-T3 as a cell surface protein involved in the transduction of activation signals. All peripheral T cells express this determinant however B cells and bone marrow cells have proven to be negative. Although the expression of this particular epitope on peripheral T cells is uniformly high, staining of thymocytes reveals distinct subpopulations of cells differing in the level of expression of this marker. This antibody will prove useful in studying the role of various components of the TCR complex in T cell activation and development, and will allow for the development of an animal model in which to investigate the immunoregulatory effects of in vivo administration of anti-CD3 antibodies, an area of obvious clinical importance. Anti-CD3e is ideal for Flow Cytometry applications, particularly as a specific marker for tracking Mouse T cells.

Purification: Protein G Affinity Chromatography.

Immunogen: H-2Kb sp from Armenian Hamster Spleen. Fusion Partner: Murine myeloma cell line SP2/0.

Clone: 145-2C11

Isotype: IgG

Application Details

Application Notes: Immunoprecipitation. Western Blot. Flow Cytometry (see Protokols).
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: 1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-M cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1x10e6 cells, representing one test). 4. To each tube add 1. 0-0. 5 µg of this antibody per 1x10e6 cells*. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (FITC Goat anti-hamster Ig) at a 1/70 dilution. PLEASENOTE: Do not use PE Goat a hamster IgG as the secondary antibody. 9. Incubate tubes at 4°C for 30-60 minutes. (It is recommended that the tubes areprotected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in Media B. 11. Resuspend the cell pellet in 50 µl ice cold Media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in phosphate buffered saline. (This stains dead cells by intercalatingDNA). MEDIA: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide (

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS containing 0.02 % Sodium Azide as preservative.

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.

Target Details for CD3

Target: CD3

Alternative Name: CD3 (CD3 Products)

Background: T cell activation through the antigen receptor (TCR) involves the cytoplasmic tails of the CD3 subunits: CD3 gamma, CD3 delta, CD3 epsilon and CD3 zeta. These CD3 subunits are structurally related members of the immunoglobulins super family encoded by closely linked genes on human chromosome 11. The CD3 components have long cytoplasmic tails that associate with cytoplasmic signal transduction molecules. This association is mediated at least in part by a double tyrosine based motif present in a single copy in the CD3 subunits. CD3 may play a role in TCR induced growth arrest, cell survival and proliferation. The CD3 antigen is present on 68-82 % of normal peripheral blood lymphocytes, 65-85 % of thymocytes and Purkinje cells in the cerebellum. It is never expressed on B or NK cells. Decreased percentages of T lymphocytes may be observed in some autoimmune diseases.Synonyms: T-cell surface antigen T3/Leu-4, T-cell surface glycoprotein CD3, T3/Leu-4

Gene ID: 12501

NCBI Accession: NP_031674

UniProt: P22646

Pathways: TCR Signaling, Ubiquitin Proteasome Pathway