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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.
Specifications

General

Verified ReactivityMouse
Antibody TypeMonoclonal
Host SpeciesRat
ImmunogenMouse thymus or spleen
Formulation0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.
Endotoxin LevelLess than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.
PreparationThe Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
ConcentrationThe antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. Older lots may have also been bottled at 1 mg/mL. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & HandlingThe antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.
ApplicationFC - Quality tested CyTOF® - Verified IHC, IP, Depletion, Block - Reported in the literature, not verified in house
Recommended UsageEach 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 million cells in 100 µl volume or 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
Application NotesClone 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, immunohistochemical staining5,6 of acetone-fixed frozen sections and zinc-fixed paraffin-embedded sections, and spatial biology (IBEX)29,30. Clone 53-6.7 is not recommended for immunohistochemistry of formalin-fixed paraffin sections. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays or in vivo studies (Cat No. 100746).
Application References(PubMed link indicates BioLegend citation)Ledbetter JA, et al. 1979. Immunol. Rev. 47:63. (IHC, IP) Hathcock KS. 1991. Current Protocols in Immunology. 3.4.1. (Deplete) Takahashi K, et al. 1992. P. Natl. Acad. Sci. USA 89:5557. (Block, IP) Ledbetter JA, et al. 1981. J. Exp. Med. 153:1503. (Block) Hata H, et al. 2004. J. Clin. Invest. 114:582. (IHC) Fan WY, et al. 2001. Exp. Biol. Med. 226:1045. (IHC) Shih FF, et al. 2006. J. Immunol. 176:3438. (FC) Kamimura D, et al. 2006. J. Immunol. 177:306. Bouwer HGA, et al. 2006. P. Natl. Acad. Sci. USA 103:5102. (FC, Deplete) Kao C, et al. 2005. Int. Immunol. 17:1607. PubMed Ko SY, et al. 2005. J. Immunol. 175:3309. (FC) PubMed Rasmussen JW, et al. 2006. Infect. Immun. 74:6590. PubMed Lee CH, et al. 2009. Clin. Cancer Res. PubMed Geiben-Lynn R, et al. 2008. Blood 112:4585. (Deplete) PubMed Kingeter LM, et al. 2008. J. Immunol. 181:6244. PubMed Guo Y, et al. 2008. Blood 112:480. PubMed Andrews DM, et al. 2008. J. Virol. 82:4931. PubMed Britschqui MR, et al. 2008. J. Immunol. 181:7681. PubMed Kenna TJ, et al. 2008. Blood 111:2091. PubMed Jordan JM, et al. 2008. Infect. Immun. 76:3717. PubMed Todd DJ, et al. 2009. J. Exp. Med. 206:2151. PubMed Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed Medyouf H, et al. 2010. Blood 115:1175. PubMed Riedl P, et al. 2009. J. Immunol. 183:370. PubMed Apte SH, et al. 2010. J. Immunol. 185:998. PubMed Bankoti J, et al. 2010. Toxicol. Sci. 115:422. (FC) PubMed del Rio ML, et al. 2011. Transpl. Int. 24:501. (FC) PubMed Cui L, et al. 2015. J Control Release. 206:220. PubMed Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product CitationsWang JX, et al. 2023. JCI Insight. 8:. PubMed Madi A, et al. 2022. Int J Cancer. 151:797. PubMed Que W, et al. 2022. Sci Adv. 8:eabo4413. PubMed Hu J, et al. 2022. J Immunother Cancer. 10:. PubMed Biram A, et al. 2020. Cell Rep. 30:1910. PubMed Vasu S, et al. 2016. Blood. 127: 2879 - 2889. PubMed Zhang D, et al. 2020. Signal Transduct Target Ther. 5:24. PubMed Yu Y, et al. 2022. Nat Commun. 13:6357. PubMed Huang Z, et al. 2021. Nat Commun. 12:145. PubMed Xu P, et al. 2020. Cancer Immunol Res. 8:1193. PubMed Wang Y, et al. 2021. Nat Commun. 12:4964. PubMed Zhang D, et al. 2020. Signal Transduct Target Ther. 5:24. PubMed Ma Q, et al. 2021. Precis Clin Med. 4:155. PubMed
RRIDAB_2810325 (BioLegend Cat. No. 100775) AB_11147171 (BioLegend Cat. No. 100746) AB_2810323 (BioLegend Cat. No. 100763) AB_2810324 (BioLegend Cat. No. 100764) AB_2810326 (BioLegend Cat. No. 100776) AB_2810327 (BioLegend Cat. No. 100777)
StructureIg superfamily, CD8α chain, 34 kD
DistributionMost thymocytes, T cell subset, some NK cells, lymphoid dendritic cells
FunctionCo-receptor for TCR
Ligand/ReceptorMHC class I molecule
Antigen References1. 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.
Gene ID12525
UniProtView information about CD8alpha on UniProt.org
Clone53-6.7
Regulatory StatusRUO
Other NamesT8, Lyt2, Ly-2
IsotypeRat IgG2a, κ