CD3 antibody

Catalog No. ABIN114132

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: 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.

Purification: Purified from ascitic fluid via Protein G Chromatography

Sterility: 0.2 μm filtered

Immunogen: H-2Kb sp

Clone: 145-2C11

Isotype: IgG

Application Details

Application Notes: This antibody will prove useful in studying the role of various components of the TCRcomplex in T cell activation and development, and will allow for the development of ananimal model in which to investigate the immunoregulatory effects of in vivoadministration of anti-CD3 antibodies, an area of obvious clinical importance. Anti-CD3 isideal for flow cytometry applications, particularly as a specific marker for tracking mouse Tcells. In addition, this monoclonal antibody, clone 145-2C11 was specifically designed totrigger T cell activation. This clone has also been reported to work in Immunoprecipitation(1, 2) and Western Blotting (Salvadori S. et al.1994. J. of Immunol.153: 5176-5182).
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: Antibody Concentration: 1 µg/10e6 cellsStrains Tested: BALB/c, C57BL/6, C3H/He, AKR/JPositive control: BALB/c, C57BL/6, C3H/He, AKR/JFlow CytometryAnalysis: 1. Prepare cell suspension in Media A. For cell preparations, deplete the redblood cell population with cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration 2x107 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1x106 cells, representing one test). 4. To each tube add 0. 2 µg of ABIN114132 per 1x106 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 100µl of secondary antibody (f. e. FITC Goat anti-hamster Ig) at a dilutionrecommended by the manufacturer. PLEASE NOTE: 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 + sodiumazide ( 100 µl of 2 M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% bovine serum albumin + sodium azide (100 µl

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS (0.2 μm filtered), with no preservative

Preservative: Without preservative

Handling Advice: Avoid repeated freezing and thawing.

Storage: -20 °C

Storage Comment: Prior to reconstitution store at -70 °C. Following reconstitution store the antibody (in aliquots) at -20 °C for 6 month.

Expiry Date: 6 months

Target Details for CD3

Target: CD3

Alternative Name: CD3 (CD3 Products)

Synonyms: CD3 antibody, AI504783 antibody, CD3epsilon antibody, T3e antibody, cd3-gamma antibody, cd3g antibody, cd3g-a antibody, cd3g/d antibody, cd3gd antibody, t3g antibody, CD3e molecule antibody, CD3d molecule antibody, CD3 antigen, epsilon polypeptide antibody, CD3g molecule S homeolog antibody, CD3E antibody, CD3D antibody, Cd3e antibody, cd3g.S antibody

Background: The e-T3 protein has been shown to be non-covalently associated on the cell surface alpha/beta 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 e-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.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