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Safety Data Sheet (SDS) Brilliant Violet 421 Anti-mouse CD3epsilon Antibody     Product Data Sheet (PDF)    
Brilliant Violet 421™ Anti-mouse CD3ε Antibody
1101675 125 µl $155.00       
1101705 50 µg $235.00       
1101680 500 µl $355.00       
Clone: 145-2C11
Isotype: Armenian Hamster IgG
Reactivity: Mouse
Immunogen: H-2Kb-specific mouse cytotoxic T lymphocyte clone BM10-37
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
Preparation: The immunoglobulin was purified by affinity chromatography and conjugated with Brilliant Violet 421™ under optimal conditions. The solution is free of unconjugated Brilliant Violet 421™ and unconjugated antibody.
Concentration: µg sizes: 0.2 mg/ml
µl sizes: lot-specific (please contact technical support for concentration and total µg amount)
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application:

FC - Quality tested
IF, IHC - Validated

Application Notes:

The 145-2C11 antibody is useful for in vitro blocking and activation assays, as well as apoptosis induction and in vivo T cell depletion. Additional reported applications (for relevant formats of this clone) include: immunoprecipitation1, immunohistochemical staining14,15 of acetone-fixed frozen sections and zinc-fixed paraffin-embedded sections, Western blotting4, complement-mediated cytotoxicity6, in vitro and in vivo stimulation of T cells1,2,7,12,16, immunofluorescent staining5, and in vivo T cell depletion8-10. The 145-2C11 antibody has been reported to block the binding of 17A2 antibody to CD3 epsilon-specific T cells11. Clone 145-2C11 is not recommended for formalin-fixed paraffin embedded sections.

Recommended Usage:

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining using the µg size, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. For immunofluorescent staining using the µl sizes, the suggested use of this reagent is ≤5 µl per million cells or 5 µl per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.

Application References:

1. Leo O, et al. 1987. P. Natl. Acad. Sci. USA 84:1374. (IP, Activ)
2. Kruisbeek AM, et al. 1991. In Current Protocols in Immunology. 3.12.1. (Activ)
3. Duke RC, et al. 1995. Current Protocols in Immunology. 3.17.1.
4. Salvadori S, et al. 1994. J. Immunol. 153:5176. (WB)
5. Payer E, et al. 1991. J. Immunol. 146:2536. (IF)
6. Jacobs H, et al. 1994. Eur. J. Immunol. 24:934. (CMCD)
7. Vossen ACTM, et al. 1995. Eur. J. Immunol. 25:1492. (Activ)
8. Henrickson M, et al. 1995. Transplantation 60:828. (Deplete)
9. Kinnaert P, et al. 1996. Transpl. Int. 9:386. (Deplete)
10. Han WR, et al. 1999. Transpl. Immunol. 7:207. (Deplete)
11. Miescher GC, et al. 1989. Immunol. Lett. 23:113. (Block)
12. Terrazas LI, et al. 2005. Intl. J. Parasitology. 35:1349. (Activ)
13. Ko SY, et al. 2005. J. Immunol. 175:3309.
14. Podd BS, et al. 2006. J. Immunol. 176:6532. (IHC)
15. Tilley SL, et al. 2007. J. Immunol. 178:3208. (IHC)
16. Wang W, et al. 2007. J. Immunol. 178:4885. (Activ)
17. Xiao S, et al. 2007. J. Exp. Med. 204:1691.
18. Chappaz S, et al. 2007. Blood doi:10.1182/blood-2007-02-074245. (FC) PubMed.
19. Curtsinger JM, et al.2005. J. Immunol. 175:4392. PubMed
20. Guo Y, et al. 2008. Blood 112:480. PubMed
21. Kenna TJ, et al. 2008. Blood 111:2091.
22. Perchonock CE, et al. 2007. J. Immunol. 179:1768. PubMed
23. Perchonock GE, et al. 2006. Mol. Cell. Biol. 26:6005. PubMed
24. Kanaya T, et al. 2008. Am. J. Physiol. Gastrointest. Liver Physiol. 295:G273. PubMed
25. de Koning BA, et al. 2006. Int. Immunol. 18:941. PubMed
26. Schulteis RD, et al. 2008. Blood 295:G273. PubMed
27. Qi Q, et al. 2009. Blood 114:564. PubMed
28. Helmersson S, et al. 2013. Am J Pathol. 9440:123. Pubmed
29. Wu S, et al. 2014. Clin Vaccine Immunol. 21:156. PubMed
30. Yan J, et al. 2014. Vaccine. 32:2833. PubMed
31. Guiterrez DA, et al. 2014. Diaebetes. 63:3827. PubMed

C57BL/6 mouse splenocytes were stained

C57BL/6 mouse splenocytes were stained with CD19 FITC and CD3ε (clone 145-2C11) Brilliant Violet 421™ (above) or Armenian hamster IgG Brilliant Violet 421™ isotype control (below).



C57BL/6 mouse splenocytes were fixed

C57BL/6 mouse splenocytes were fixed with 2% paraformaldehyde (PFA), and then stained with 5 µg/ml CD3 (clone 145-2C11) Brilliant Violet 421™ (blue) and 5 µg/ml CD19 (clone 6D5) Alexa Fluor® 647 (red) for 30 minutes at room temperature. The image was captured by 40X objective.



Description:

CD3ε is a 20 kD transmembrane protein, also known as CD3 or T3. It 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. CD3ε forms a TCR complex by associating with the CD3δ, γ and ζ chains, as well as the 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.

Other Names: CD3ε, T3, CD3
Structure: Ig superfamily, forms CD3/TCR complex with CD3δ, γ and ζ subunits and TCR (α/β and γ/δ), 20 kD
Distribution: Thymocytes (differentiation dependent), mature T cells, NK-T cells
Function: TCR signal transduction, T cell activation, antigen recognition
Ligand Receptor: Peptide antigen/MHC-complex
Antigen References:

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Davis MM. 1990. Annu. Rev. Biochem. 59:475.
3. Weiss A, et al. 1994. Cell 76:263.