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Multiparameter flow cytometric analysis of CD8 expression on human peripheral blood leukocyte populations. Human whole blood was stained with either APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; Left Plot) or APC Mouse Anti-Human CD8 antibody (Cat. No. 570816/570892; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD8 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
BD Pharmingen™ APC Mouse Anti-Human CD8
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The SK1 monoclonal antibody specifically binds to CD8 alpha (CD8α). CD8α is a type I transmembrane glycoprotein and a member of the immunoglobulin superfamily. CD8α is expressed by the majority of thymocytes, by subpopulations of αβ T cells and γδ T cells and by some NK cells. Cell surface CD8α is expressed either as a disulfide-linked homodimer (CD8αα) or as a heterodimer (CD8αβ) when disulfide-bonded to a CD8 beta chain (CD8β). CD8-positive αβ T cells coexpress both CD8αα homodimers and CD8αβ heterodimers whereas some γδ T cells and NK cells express CD8αα homodimers. CD8 plays important roles in T cell activation and selection. The extracellular IgSF domain of CD8α binds to a non-polymorphic determinant on HLA class I molecules (α3 domain) and enables CD8 to function as a co-receptor with MHC class I-restricted TCR during T cell recognition of antigen. The cytoplasmic domain of CD8α associates with Lck, a Src family protein tyrosine kinase that is involved in intracellular signaling.
Development References (13)
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Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
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Bernard A, Boumsell L, Hill C. Joint report of the first international workshop on human leucocyte differentiation antigens by the investigators of the participating laboratories: T2 protocol. In: Bernard A. A. Bernard .. et al., ed. Leucocyte typing : human leucocyte differentiation antigens detected by monoclonal antibodies : specification, classification, nomenclature = Typage leucocytaire : antigènes de différenciation leucocytaire humains révélés par les anticorps monoclonaux : "Rapports des études communes". Berlin New York: Springer-Verlag; 1984:25-60.
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Dongworth DW, Gotch FM, Carter NP, Hildreth PDK, McMichael AJ. Inhibition of virus-specific, HLA-restricted, T cell-mediated lysis by monoclonal anti-T cell antibodies. In: Bernard A. A. Bernard .. et al., ed. Leucocyte typing : human leucocyte differentiation antigens detected by monoclonal antibodies : specification, classification, nomenclature = Typage leucocytaire : antigènes de différenciation leucocytaire humains révélés par les anticorps monoclonaux : "Rapports des études communes". Berlin New York: Springer-Verlag; 1984:320-328.
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Engleman EG, Benike CJ, Glickman E, Evans RL. Antibodies to membrane structures that distinguish suppressor/cytotoxic and helper T lymphocyte subpopulations block the mixed leukocyte reaction in man. J Exp Med. 1981; 154(1):193-198. (Clone-specific: Cell separation, Flow cytometry, Functional assay, Inhibition). View Reference
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Engleman EG, Benike CJ, Grumet FC, Evans RL. Activation of human T lymphocyte subsets: helper and suppressor/cytotoxic T cells recognize and respond to distinct histocompatibility antigens. J Immunol. 1981; 127(5):2124-2129. (Clone-specific: Cell separation, Flow cytometry, Fluorescence activated cell sorting). View Reference
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Evans RL, Wall DW, Platsoucas CD, et al. Thymus-dependent membrane antigens in man: inhibition of cell-mediated lympholysis by monoclonal antibodies to TH2 antigen. Proc Natl Acad Sci U S A. 1981; 78(1):544-548. (Immunogen: Flow cytometry, Functional assay, Inhibition). View Reference
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Jonker M, Meurs G. Monoclonal antibodies specific for B cells, cytotoxic/suppressor T cells, and a subset of cytotoxic/suppressor T cells in the Rhesus monkey. In: Bernard A. A. Bernard .. et al., ed. Leucocyte typing : human leucocyte differentiation antigens detected by monoclonal antibodies : specification, classification, nomenclature = Typage leucocytaire : antigènes de différenciation leucocytaire humains révélés par les anticorps monoclonaux : "Rapports des études communes". Berlin New York: Springer-Verlag; 1984:328-336.
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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Ledbetter JA, Evans RL, Lipinski M, Cunningham-Rundles C, Good RA, Herzenberg LA. Evolutionary conservation of surface molecules that distinguish T lymphocyte helper/inducer and cytotoxic/suppressor subpopulations in mouse and man. J Exp Med. 1981; 153(2):310-323. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
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McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:1-1050.
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Reichert T, DeBruyere M, Deneys V, et al. Lymphocyte subset reference ranges in adult Caucasians. Clin Immunol Immunopathol. 1991; 60(2):190-208. (Biology). View Reference
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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Warner NL, Lanier LL, Jackson A, Babcock G, Evans R. Multiparameter approaches to FACS analysis of human leucocyte cell surface antigens. In: Bernard A. A. Bernard .. et al., ed. Leucocyte typing : human leucocyte differentiation antigens detected by monoclonal antibodies. Berlin New York: Springer-Verlag; 1984:621-630.
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.