The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Application:
FC
Recommended Usage:
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤ 0.5 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome. ** Pacific Blue™ is a registered trademark of Molecular Probes, Inc. Pacific Blue™ dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
Application Notes:
Additional reported applications (for the relevant formats) include: immunoprecipitation1, complement-mediated cytotoxicity1,3, immunohistochemical staining1,4 of acetone-fixed frozen sections, in vitro stimulation of T cells1 and depletion2 of CD3+ cells in vivo. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 100208).
Application References:
1. Miescher, G.C., et al., 1989. Immunol. Lett. 23:113. 2. Mysliwietz, J., et al., 1992. Blood 80:2661. 3. Wu, L., et al., 1991. J. Exp. Med. 174:1617. 4. Zhang, Y., et al., 2002. J. Immunol. 168: 3088. 5. Zan, H., et al., 2005. EMBO J. 24: 3757. 6. Achtman, AH., et al. 2009. J. Leukoc Biol. 85:409. PubMed 7. Chiossone, L., et al. 2009. Blood. 113:5488. PubMed
C57BL/6 mouse splenocytes stained with Pacific Blue™ 17A2
Description:
CD3, also known as T3, is a member of the Ig superfamily and primarily expressed on T cells, NK-T cells, and at different levels on thymocytes during T cell differentiation. CD3 is composed of CD3ε, δ, γ and ζ chains. It forms a TCR complex by associating with TCR α/β or γ/δ chains. CD3 plays a critical role in TCR signal transduction, T cell activation, and antigen recognition by binding the peptide/MHC antigen complex. The 17A2 antibody recognizes ε/γ (but not ε/δ) of CD3 complex. The 17A2 antibody can induce T cell activation and has been reported to deplete CD3+ cells in vivo.
Other Names:
T cell antigen receptor complex, T3
Structure:
Ig superfamily, CD3/TCR, 20 kD
Distribution:
Thymocytes (differentiation dependent), mature T cells, NK-T cells
Function:
Antigen recognition, TCR signal transduction, T cell activation
Ligand Receptor:
Peptide antigen/MHC-complex
Antigen References:
1. Barclay, A., et al., 1997. The Leukocyte Antigen FactsBook, Academic Press. 2. Davis, M.M. 1990. Annu. Rev. Biochem. 59:475. 3. Weiss, A. et al., 1994. Cell 76:263.
