Phosphate-buffered solution, pH 7.2, containing no preservative. 0.2 μm filter sterilized. Endotoxin level is < 0.1 EU/μg of the protein (< 0.01 ng/μg of the protein) as determined by the LAL test.
Preparation:
The LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Storage & Handling:
The antibody solution should be stored undiluted at 4 °C. This LEAF™ solution contains no preservative; handle under aseptic conditions.
Application:
FC - Quality tested IHC, IP, Depletion, Blocking - Reported in the literature
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 ≤ 0.25 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
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, zinc-fixed paraffin-embedded sections and 10% formalin-fixed paraffin-embedded sections6. Special Note: The antibody works well on acetone-fixed frozen sections as well as Zinc-fixed paraffin-embedded sections. It sometime works on formalin-fixed paraffin-embedded tissue sections but inconsistent results have been reported. The 53-6.7 antibody competes with 5H10-1 antibody (Cat. No. 100802) for binding to thymocytes3. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 100716).
Application References:
1. Ledbetter, J.A., et al. 1979. Immunol. Rev. 47:63. (IHC,IP) 2. Hathcock, K.S. 1991. Current Protocols in Immunology. 3.4.1. (Deplete) 3. Takahashi, K., et al. 1992. PNAS 89:5557. (Block, IP) 4. Ledbetter, J.A., et al. 1981. J. Exp. Med. 153:1503. (Block) 5. Hata, H., et al. 2004. J. Clin. Invest. 114:582. (IHC) 6. Fan, W.Y., et al. 2001. Exp. Biol. Med. 226:1045. (IHC) 7. Ko, S.Y., et al. 2005. J. Immunol. 175:3309. (IHC) 8. Shih, F.F., et al. 2006. J. Immunol. 176:3438. (FC) 9. Kamimura, D., et al. 2006. J. Immunol. 177: 306. 10. Bouwer, H.G.A., et al. 2006. PNAS 103:5102. (FC, Deplete) 11. Kao, C., et al. 2005. Int. Immunol. 17:1607. PubMed 12. Ko, S.Y., et al. 2005. J. Immunol. 175:3309. PubMed 13. Rasmussen, J.W., et al. 2006. Infect. Immune. 74:6590. PubMed 14. Lee, CH., et al. 2009. Clin. Cancer Res. PubMed 15. Geiben-Lynn, R., et al., 2008. Blood 112:4585. (Deplete) PubMed 16. Kingeter, LM., et al. 2008. J. Immunol. 181:6244. PubMed 17. Guo, Y., et al. 2008. Blood. 112:480. PubMed. 18. Andrews, D.M., et al. 2008. J. Virol. 82:4931. PubMed 19. Britschqui, M.R., et al. 2008. J. Immunol. 181:7681. PubMed 20. Kenna, T.J., et al. 2008. Blood. 111:2091. PubMed 21. Jordan, J.M., et al.2008. Infect. Immun. 76:3717. PubMed 22. Todd, DJ., et al. 2009. J. Exp Med. PubMed 23. Bankoti, J., et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed 24. Medyouf, H., et al. 2010. Blood. 115:1175. PubMed 25. Riedl, P., et al. 2009. J. Immunol. 183:370. PubMed 26. Apte, S.H., et al. 2010. J. Immunol. 185:998. PubMed
C57BL/6 mouse splenocytes stained with LEAF™ purified 53-6.7, followed by anti-rat IgGs-FITC
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 34kD 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. 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.
Other Names:
T8, Lyt2, Ly-2
Structure:
Ig superfamily, CD8α chain, 34 kD
Distribution:
Most thymocytes, T cell subset, some NK 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.
