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Safety Data Sheet (SDS) Purified Anti-mouse CD8a Antibody     Product Data Sheet (PDF)    
Purified Anti-mouse CD8a Antibody
1103505 50 µg $35.00       
1103510 500 µg $95.00       
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.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application:

FC - Quality tested

IP - Reported in the literature

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 flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. For immunohistochemical staining on formalin-fixed paraffin-embedded tissue sections, the suggested use of this reagent is 5.0 - 10 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

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. Cui L, et al. 2015. J Control Release. 206:220. PubMed

C57BL/6 mouse splenocytes were stained

C57BL/6 mouse splenocytes were stained with purified CD8 (clone 53-6.7) (filled histogram) or rat IgG2a, κ isotype control (open histogram), followed by anti-rat IgG FITC.

C57BL/6 frozen mouse spleen section

C57BL/6 frozen mouse spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS for 30 minutes at room temperature. Then the section was stained with 10 µg/mL of purified anti-mouse CD8a (clone 53-6.7), 10 µg/mL of Alexa Fluor® 647 anti-CD20 (green) overnight at 4°C, followed by 2.5 µg/mL of Alexa Fluor® 594 anti-rat IgG2a (clone MRG2a-83) (red) for two hours at room temperature. The image was captured by 10X objective.



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.