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 antibody was purified by affinity chromatography.
Concentration:
1 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4 °C. This LEAF™ solution contains no preservative; handle under aseptic conditions.
Application:
FC8-10 - Quality Tested IHC(Frozen/ Paraffin)1,6,7*, IP2,4*, Depletion3,11*, Blocking4,5 * - *This application has been 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: immunoprecipitation2,4, in vivo and in vitro cell depletion3,11, inhibition of CD8 T cell proliferation4, blocking of cytotoxicity4,5, and immunohistochemical staining1,6,7 of acetone-fixed frozen sections, zinc-fixed paraffin-embedded sections and 10% formalin-fixed paraffin-embedded sections7. 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 thymocytes4. 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. Nitta, H., et al., 1997. Cell Vision 4:73. 2. Ledbetter, J.A., et al., 1979. Immunol. Rev. 47:63. 3. Hathcock, K.S. 1991. Current Protocols in Immunology. pp. 3.4.1 - 3.4.3. 4. Takahashi, K., et al., 1992. PNAS 89:5557. 5. Ledbetter, J.A., et al., 1981. J. Exp. Med. 153:1503. 6. Hata, H., et al., 2004. J. Clin. Invest. 114:582. 7. Fan, W. Y., et al., 2001. Exp. Biol. Med. 226: 1045. 8. Ko, S-Y., et al., 2005. J. Immunol. 175:3309. 9. Shih, F. F., et al., 2006. J. Immunol. 176:3438. 10. Kamimura, D., et al., 2006. J. Immunol. 177: 306. 11. Bouwer, H.G.A., et al., 2006. PNAS 103:5102.PubMed 12. Jones, TG., et al. 2009. J. Immunol. 183:5251. 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.
