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Cat. No |
Model |
Description |
Unit |
Price (VATº°µµ) |
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Àç°í |
DataSheet |
ÁÖ¹®/°ßÀû |
LCA-24-66135 |
CL001A |
Hamster Anti-CD3e (T3) Monoclonal Antibody, Unconjugated, Clone 145-2C11/1 mg/Mouse/Hamster |
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Subject
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Hamster Anti-CD3e (T3) Monoclonal Antibody, Unconjugated, Clone 145-2C11 |
Description
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1 mg / Mouse/Hamster |
Clonality
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Mono |
Company
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Acris Antibodies GmbH |
Application
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Frozen sections, Flow cytometry, Western blot / Immunoblot, Functional assay, Immunoprecipitation |
Conjugation
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Unconjugated |
Immunogen
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CD3e (T3) |
Contents
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Description: Manufacturer | Acris Antibodies GmbH | Quantity | 1 mg | Reactivity | Mouse | Presentation | Azide Free | Applications | Frozen sections, Flow cytometry, Western blot / Immunoblot, Functional assay, Immunoprecipitation | Isotype | IgG | Host | Hamster | Clone | 145-2C11 | PDF datasheet | view | Shipping to | Worldwide | Synonyms | T3/Leu-4 | Swiss Prot Num | P22646 | Immunogen | H-2Kb sp | Product Type | Antibodies | Storage | Store at 4°C. For long term storage, aliquot and freeze unused portions at -20°C in volumes appropriate for single usage. Avoid repeated freeze/thaw cycles. Should be handled under aseptic conditions. | References | 1. Leo, O., Foo. M., Sachs, D.H. Samelson, L.E. and J.A. Bluestone. 1987. Identification of a monoclonal antibody specific for a murine TS polypeptide. Proc. Natl Acad. Sci. USA 84: 1374 - 1378. 2. Portoles, P., Rojo, J., Golby, A., Bonneville, M,. Gromkowski, S., Greenbaum, L., Janeway, G., Murphy, D., and K. Bottomly. Monoclonal Antibodies to Murine CD3u? Define Distinct Epitopes, One of which may interact with CD4 During T Cell Activation. J. of Immunology 142: 4168 - 4175. 1989. 3. Bluestone, J.A., Pardoll, D., Sharrow, S.O. and B.J. Fowlkes. 1987. Characterization of murine thymocytes with CD3 - associated T cell receptor structure Nature. 326, 82 - 84. 4. Hirsch, R., Gress, R.E., Pluznik, D.H., Eckhaus, M. and J.A. Bluestone. 1989. Effects of in vitro Administration of Anti-CD3 Monoclonal Antibody on T Cell Function in Mice II. In Vitro Activation of T Cells. J. of Immunology 142: 737-743. 5. Hirsch, R., Eckhaus, M., Auchincloss, H., Jr., Sachs, D.H. and J.A. Bluestone. 1988. Effects of in vivo Administration of Anti-T3 Monoclonal Antibody on T Cell Function in Mice. J. of Immun | Protocols | Antibody Concentration: 1 ug/10e6 cells Strains Tested: BALB/c, C57BL/6, C3H/He, AKR/J Positive control: BALB/c, C57BL/6, C3H/He, AKR/J Flow Cytometry Analysis: 1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood 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 ul of this suspension to each tube (each tube will then contain 1x106 cells, representing one test). 4. To each tube add 0.2 ug of CL001A per 1x106 cells*. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4?¡ÆC. 7. Wash 2 times at 4?¡ÆC. 8. Add 100ul of secondary antibody (f.e. FITC Goat anti-hamster Ig) at a dilution recommended by the manufacturer. PLEASE NOTE: Do not use PE Goat a hamster IgG as the secondary antibody. 9. Incubate tubes at 4?¡ÆC for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4?¡ÆC in Media B. 11. Resuspend the cell pellet in 50 ul ice cold Media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 ul of propidium iodide at 0.5 mg/ml in phosphate buffered saline. (This stains dead cells by intercalating DNA). Media: A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide ( 100 ul of 2 M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7.2) + 0.5% bovine serum albumin + sodium azide (100 ul of 2 M sodium azide in 100 mls). | |
Format | State: Liquid | ? | Purification: Purified from ascitic fluid via Protein G Chromatography | ? | Buffer System: PBS (0.2 um filtered), with no preservative | |
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. | Application | 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-CD3 is ideal for flow cytometry applications, particularly as a specific marker for tracking mouse T cells. In addition, this monoclonal antibody, clone 145-2C11 was specifically designed to trigger 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). | 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. | Concentration | 1.0 mg/ml | |
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