Biolegend, 100746, Ultra-LEAF™ Purified anti-mouse CD8a Antibody, 1mg

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.
1mg
Verified Reactivity: Mouse
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: Mouse thymus or spleen
Formulation: 0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.
Endotoxin Level: Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.
Preparation: The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration: The 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 & Handling: The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.
Application: FC - Quality tested CyTOF® - Verified IHC, IP, Depletion, Block - Reported in the literature, not verified in house
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 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 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, 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 Citations: Wang 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
RRID: AB_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)
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.
Gene ID: 12525
UniProt: View information about CD8alpha on UniProt.org
Clone: 53-6.7
Regulatory Status: RUO
Other Names: T8, Lyt2, Ly-2
Isotype: Rat IgG2a, κ
Q: Do you guarantee that your antibodies are totally pathogen free?
A: BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.
Q: Does BioLegend test each Ultra-LEAF™ antibody by functional assay?
A: No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.
Q: Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?
A: No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.
Q: Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?
A: We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.