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Safety Data Sheet (SDS) Brilliant Violet 421 Anti-mouse CD8a Antibody     Product Data Sheet (PDF)    
Brilliant Violet 421™ Anti-mouse CD8a Antibody
1103685 125 µl $150.00       
1103765 50 µg $235.00       
1103690 500 µl $345.00       
Clone: 53-6.7
Isotype: Rat IgG2a, κ
Reactivity: Mouse
Immunogen: Mouse thymus or spleen
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).
Preparation: The antibody 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
IHC - Validated

Application Notes:

Clone 53-6.7 antibody competes with clone 5H10-1 antibody for binding to thymocytes3. The 53-6.7 antibody has been reported to block antigen presentation via MHC class I and inhibit T cell responses to IL-2. This antibody has also been used for depletion of CD8a+ cells. Additional reported applications (for the relevant formats) include: immunoprecipitation1,3, in vivo and in vitro cell depletion2,10,15, inhibition of CD8 T cell proliferation3, blocking of cytotoxicity3,4, and immunohistochemical staining5,6 of acetone-fixed frozen sections and zinc-fixed paraffin-embedded sections. Clone 53-6.7 is not recommended for immunohistochemistry of formalin-fixed paraffin 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 ≤0.5 µg per million cells in 100 µl volume. For immunofluorescent staining using 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. Ledbetter JA, et al. 1979. Immunol. Rev. 47:63. (IHC, IP)
2. Hathcock KS. 1991. Current Protocols in Immunology. 3.4.1. (Deplete)
3. Takahashi K, et al. 1992. P. Natl. Acad. Sci. USA 89:5557. (Block, IP)
4. Ledbetter JA, et al. 1981. J. Exp. Med. 153:1503. (Block)
5. Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC)
6. Fan WY, et al. 2001. Exp. Biol. Med. 226:1045. (IHC)
7. Shih FF, et al. 2006. J. Immunol. 176:3438. (FC)
8. Kamimura D, et al. 2006. J. Immunol. 177:306.
9. Bouwer HGA, et al. 2006. P. Natl. Acad. Sci. USA 103:5102. (FC, Deplete)
10. Kao C, et al. 2005. Int. Immunol. 17:1607. PubMed
11. Ko SY, et al. 2005. J. Immunol. 175:3309. (FC) PubMed
12. Rasmussen JW, et al. 2006. Infect. Immun. 74:6590. PubMed
13. Lee CH, et al. 2009. Clin. Cancer Res. PubMed
14. Geiben-Lynn R, et al. 2008. Blood 112:4585. (Deplete) PubMed
15. Kingeter LM, et al. 2008. J. Immunol. 181:6244. PubMed
16. Guo Y, et al. 2008. Blood 112:480. PubMed
17. Andrews DM, et al. 2008. J. Virol. 82:4931. PubMed
18. Britschqui MR, et al. 2008. J. Immunol. 181:7681. PubMed
19. Kenna TJ, et al. 2008. Blood 111:2091. PubMed
20. Jordan JM, et al. 2008. Infect. Immun. 76:3717. PubMed
21. Todd DJ, et al. 2009. J. Exp. Med. 206:2151. PubMed
22. Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed
23. Medyouf H, et al. 2010. Blood 115:1175. PubMed
24. Riedl P, et al. 2009. J. Immunol. 183:370. PubMed
25. Apte SH, et al. 2010. J. Immunol. 185:998. PubMed
26. Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed
27. del Rio ML, et al. 2011. Transpl. Int. 24:501. (FC) PubMed
28. Chen Z, et al. 2014. Cancer Immunol Res. 2:911. PubMed
29. Ballet R, et al. 2014. PLoS Pathog. 10:1004550. PubMed
30. Hu-Lieskovan S, et al. 2015. Sci Transl Med. 7:279. PubMed

C57BL/6 mouse splenocytes were stained

C57BL/6 mouse splenocytes were stained with CD3ε FITC and CD8a (clone 53-6.7) Brilliant Violet 421™. Quadrant gating was based on the rat IgG2a, κ Brilliant Violet 421™ isotype control.

BL6 mouse lymph nodes, fixed

BL6 mouse lymph nodes, fixed O/N in PLP, blocked with 10% rat serum, stained with CD8a-BV421 (red), B220-Alexa Fluor® 647 (blue), and TCRβ-Alexa Fluor® 488 (green) in 1% BSA and 0.1% Tween-20 in PBS. Images were acquired with an automated widefield microscope (Nikon Eclipse Ti) and a CCD camera (QImaging Retiga 2000R). Emitted light was collected through 440/40, 525/50, and 700/75 nm bandpass filters. Images provided by Ann Haberman and Christine Podolski, Yale University.



Description:

CD8, also known as Lyt-2, Ly-2, or T8, consists of disulfide-linked α and β chains that form the α(CD8a)/β(CD8b) heterodimer and α/α homodimer. CD8a is a 34 kD protein that belongs to the immunoglobulin family. The CD8 α/β heterodimer is expressed on the surface of most thymocytes and a subset of mature TCR α/β T cells. CD8 expression on mature T cells is non-overlapping with CD4. The CD8 α/α homodimer is expressed on a subset of γ/δ TCR-bearing T cells, NK cells, intestinal intraepithelial lymphocytes, and lymphoid dendritic cells. CD8 is an antigen co-receptor on T cells that interacts with MHC class I on antigen-presenting cells or epithelial cells. CD8 promotes T cell activation through its association with the TCR complex and protein tyrosine kinase lck.

Other Names: T8, Lyt2, Ly-2
Structure: Ig superfamily, CD8α chain, 34 kD
Distribution: Most thymocytes, T cell subset, some NK cells, lymphoid dendritic cells
Function: Co-receptor for TCR
Ligand Receptor: MHC class I molecule
Antigen References:

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Zamoyska R. 1994. Immunity 1:243.
3. Ellmeier W, et al. 1999. Annu. Rev. Immunol. 17:523.