Multiparameter flow cytometric analysis of CD8 expression on Human peripheral blood leucocyte populations. Fresh whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426; Top Plots) and with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949; Left Plots) or BD Horizon™ R718 Mouse Anti-Human CD8 antibody (Cat. No. 569415/569416; Right Plots). Bivariate pseudocolor density plots showing the correlated expression of CD8 (or Ig Isotype control staining) versus CD3 (Top Plots) or side-light scatter (SSC-A) signals (Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Multiparameter flow cytometric analysis of CD8 expression on Human peripheral blood leucocyte populations. Fresh whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426; Top Plots) and with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949; Left Plots) or BD Horizon™ R718 Mouse Anti-Human CD8 antibody (Cat. No. 569415/569416; Right Plots). Bivariate pseudocolor density plots showing the correlated expression of CD8 (or Ig Isotype control staining) versus CD3 (Top Plots) or side-light scatter (SSC-A) signals (Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
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Multiparameter flow cytometric analysis of CD8 expression on Human peripheral blood leucocyte populations. Fresh whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426; Top Plots) and with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949; Left Plots) or BD Horizon™ R718 Mouse Anti-Human CD8 antibody (Cat. No. 569415/569416; Right Plots). Bivariate pseudocolor density plots showing the correlated expression of CD8 (or Ig Isotype control staining) versus CD3 (Top Plots) or side-light scatter (SSC-A) signals (Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Multiparameter flow cytometric analysis of CD8 expression on Human peripheral blood leucocyte populations. Fresh whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leucocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426; Top Plots) and with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949; Left Plots) or BD Horizon™ R718 Mouse Anti-Human CD8 antibody (Cat. No. 569415/569416; Right Plots). Bivariate pseudocolor density plots showing the correlated expression of CD8 (or Ig Isotype control staining) versus CD3 (Top Plots) or side-light scatter (SSC-A) signals (Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Product Details
BD Horizon™
CD8; CD8A; Leu2; OKT8 T-cell antigen; T8 T-cell antigen
Human (QC Testing), Rhesus,Cynomolgus,Baboon (Reported)
Mouse BALB/c x A/J, also known as CAF1 IgG2a, κ
Normal Human Thymocytes
Flow cytometry (Routinely Tested)
5 µl/test
HLDA VI: T 6T-CD8.3; T 6T-R8; BP 495
925
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.
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 cells and BD CompBead 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.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Human BD Fc Block™ RUO
Size
0.25 mg
Cat No.
564220
BV421 Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
562426
569416 Rev. 1
Antibody Details
OKT8
The OKT8 monoclonal antibody specifically recognizes CD8 alpha (CD8a or CD8α). CD8a is a ~32-34 kDa type I transmembrane glycoprotein that is encoded by CD8A, which belongs to the immunoglobulin superfamily (IgSF), and is commonly referred to as CD8. CD8a is expressed by the majority of thymocytes, subpopulations of αβ T cells and γδ T cells, and by subsets of NK cells. Cell surface CD8a is expressed either as a disulfide-linked homodimer (CD8αα) or as a heterodimer (CD8αβ) when disulfide-bonded to a CD8 beta chain (CD8b or 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 coreceptor 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. The OKT8 antibody reportedly crossreacts with CD8a expressed by leucocytes from some non-human primate (NHP) species.
569416 Rev. 1
Format Details
R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
R718
Red 627-640 nm
695 nm
718 nm
569416 Rev.1
Citations & References
View product citations for antibody "569416" on CiteAb
Development References (8)
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Hoffman RA, Kung PC, Hansen WP, Goldstein G.. Simple and rapid measurement of human T lymphocytes and their subclasses in peripheral blood.. Proc Natl Acad Sci U S A. 1980; 77(8):4914-4917. (Clone-specific: Flow cytometry). View Reference
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Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
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Mattila JT, Diedrich CR, Lin PL, Phuah J, Flynn JL. Simian immunodeficiency virus-induced changes in T cell cytokine responses in cynomolgus macaques with latent Mycobacterium tuberculosis infection are associated with timing of reactivation.. J Immunol. 2011; 186(6):3527-37. (Clone-specific: Flow cytometry). View Reference
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Myburgh JA, Smit JA, Stark JH, Browde S. Total lymphoid irradiation in kidney and liver transplantation in the baboon: prolonged graft survival and alterations in T cell subsets with low cumulative dose regimens.. J Immunol. 1984; 132(2):1019-25. (Clone-specific: Cytotoxicity, Depletion). View Reference
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Perussia B, Fanning V, Trinchieri G. A human NK and K cell subset shares with cytotoxic T cells expression of the antigen recognized by antibody OKT8.. J Immunol. 1983; 131(1):223-31. (Clone-specific: Flow cytometry). View Reference
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Reimann KA, Waite BC, Lee-Parritz DE, et al. Use of human leukocyte-specific monoclonal antibodies for clinically immunophenotyping lymphocytes of rhesus monkeys. Cytometry. 1994; 17(1):102-108. (Clone-specific: Flow cytometry). View Reference
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Reinherz EL. Reinherz EL, Haynes BF, Nadler LM, Bernstein ID, ed. Leukocyte Typing II. New York, NY: Springer-Verlag; 1986:1-549.
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Shiue L, Gorman SD, Parnes JR. A second chain of human CD8 is expressed on peripheral blood lymphocytes.. J Exp Med. 1988; 168(6):1993-2005. (Clone-specific: Flow cytometry). View Reference
569416 Rev. 1
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.