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The CD8a antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration:
0.5 mg/ml
Storage & Handling:
The CD8a antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
This CD8a reagent is developed for immunofluorescent staining for flow cytometric analysis; the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. It is highly recommended that the reagent be titrated for optimal performance for each application.
* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome. ** Pacific Blue™ is a registered trademark of Molecular Probes, Inc. Pacific Blue™ dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
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, zinc-fixed paraffin-embedded sections. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 100716). For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 100746) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin <0.01 EU/µg). Clone 53-6.7 antibody competes with 5H10-1 antibody (Cat. No. 100802) for binding to thymocytes3.
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. Ko SY, et al. 2005. J. Immunol. 175:3309. (IHC) 8. Shih FF, et al. 2006. J. Immunol. 176:3438. (FC) 9. Kamimura D, et al. 2006. J. Immunol. 177:306. 10. Bouwer HGA, et al. 2006. P. Natl. Acad. Sci. USA 103:5102. (FC, Deplete) 11. Kao C, et al. 2005. Int. Immunol. 17:1607. PubMed 12. Ko SY, et al. 2005. J. Immunol. 175:3309. PubMed 13. Rasmussen JW, et al. 2006. Infect. Immun. 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 DM, et al. 2008. J. Virol. 82:4931. PubMed 19. Britschqui MR, et al. 2008. J. Immunol. 181:7681. PubMed 20. Kenna TJ, et al. 2008. Blood 111:2091. PubMed 21. Jordan JM, et al. 2008. Infect. Immun. 76:3717. PubMed 22. Todd DJ, et al. 2009. J. Exp. Med. 206:2151. 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 SH, et al. 2010. J. Immunol. 185:998. PubMed 27. Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) Pacific Blue™ (filled histogram) or rat IgG2a, κ Pacific Blue™ isotype control (open histogram).
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.
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
APC anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) APC (filled histogram) or rat IgG2a, κ APC isotype control (open histogram).
Biotin anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with biotinylated CD8 (clone 53-6.7) (filled histogram) or rat IgG2a, κ isotype control (open histogram), followed by Sav-PE.
FITC anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) FITC (filled histogram) or rat IgG2a, κ FITC isotype control (open histogram).
LEAF™ Purified anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with LEAF™ purified CD8 (clone 53-6.7) (filled histogram) or rat IgG2a, κ isotype control (open histogram), followed by anti-rat IgG FITC.
PE anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) PE (filled histogram) or rat IgG2a, κ PE isotype control (open histogram).
PE/Cy5 anti-mouse CD8a
C57BL/6 splenocytes were stained with CD8 (clone 53-6.7) PE/Cy5 and CD3 FITC.
Purified anti-mouse CD8a
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.
PE/Cy7 anti-mouse CD8a
C57BL/6 splenocytes were stained with CD8 (clone 53-6.7) PE/Cy7 and CD3 FITC.
APC/Cy5.5 anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) APC/Cy5.5 (filled histogram) or rat IgG2a, κ APC/Cy5.5 isotype control (open histogram).
APC/Cy7 anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) APC/Cy7 (filled histogram) or rat IgG2a, κ APC/Cy7 isotype control (open histogram).
Alexa Fluor® 488 anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) Alexa Fluor® 488 (solid line) or rat IgG2a, κ Alexa Fluor® 488 isotype control (broken line).
Alexa Fluor® 647 anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) Alexa Fluor® 647 (filled histogram) or rat IgG2a, κ Alexa Fluor® 647 isotype control (open histogram).
Pacific Blue™ anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) Pacific Blue™ (filled histogram) or rat IgG2a, κ Pacific Blue™ isotype control (open histogram).
Alexa Fluor® 700 anti-mouse CD8a
C57BL/6 mouse splenocytes stained with CD8 (clone 53-6.7) Alexa Fluor® 700 (filled histogram) or rat IgG2a, κ Alexa Fluor® 700 isotype control (open histogram).
PerCP/Cy5.5 anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD8 (clone 53-6.7) PerCP/Cy5.5 (filled histogram) or rat IgG2a, κ PerCP/Cy5.5 isotype control (open histogram).
PerCP anti-mouse CD8a
C57BL/6 thymocytes were stained with CD8 (clone 53-6.7) PerCP and CD4 APC.
Brilliant Violet 421™ anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD3ε FITC and CD8a (clone 53-6.7) Brilliant Violet 421™ (top) or rat IgG2a, κ Brilliant Violet 421™ isotype control (bottom).
Brilliant Violet 570™ anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD3 FITC and CD8a (clone 53-6.7) Brilliant Violet 570™ (top) or rat IgG2a, κ Brilliant Violet 570™ isotype control (bottom).
Brilliant Violet 650™ anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with CD3 PE and CD8a (clone 53-6.7) Brilliant Violet 650™.
Ultra-LEAF™ Purified anti-mouse CD8a
C57BL/6 mouse splenocytes were stained with LEAF™ purified CD8 (clone 53-6.7) (filled histogram) or rat IgG2a, κ isotype control (open histogram), followed by anti-rat IgG FITC.
