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V500 Mouse Anti-Human CD8
V500 Mouse Anti-Human CD8

Flow cytometric analysis of CD8 on human lymphocytes.  Whole blood was stained with BD Horizon™ V500 Mouse Anti-Human CD8 (Cat. No. 560775/560774; solid line histogram) and compared to whole blood stained with BD Horizon™ V500 Mouse IgG1, κ Isotype Control (Cat. No. 560787; dashed line histogram). Lymphocytes were selected by light scatter profile. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.

Flow cytometric analysis of CD8 on human lymphocytes.  Whole blood was stained with BD Horizon™ V500 Mouse Anti-Human CD8 (Cat. No. 560775/560774; solid line histogram) and compared to whole blood stained with BD Horizon™ V500 Mouse IgG1, κ Isotype Control (Cat. No. 560787; dashed line histogram). Lymphocytes were selected by light scatter profile. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.

Product Details
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BD Horizon™
CD8α; CD8A; CD8 alpha; Leu2; MAL; T8; p32
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse IgG1, κ
Human CD8a
Flow cytometry (Routinely Tested)
5 µl
IV T171; V T-CD08.03; VI 6T-CD8.1, 6T-081
925
AB_1937333
Aqueous buffered solution containing protein stabilizer, glycerol and ≤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 with BD Horizon™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon V500 is covered by one or more of the following US patents: 8,431,416.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560774 Rev. 2
Antibody Details
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RPA-T8

The RPA-T8 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. The RPA-T8 and HIT8a monoclonal antibodies are not cross-blocking.  This clone has been reported to react with a subset of peripheral blood lymphocytes, but not monocytes nor granuloyctes, of baboon and both rhesus and cynomolgus macaque monkey. In general, a higher frequency of CD8+ and CD4+CD8+ lymphocytes are observed in non-human primates compared to normal human donors.

The antibody is conjugated to BD Horizon V500, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.

When compensating dyes in this spectral range (such as BD Horizon V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls.  Due to spectral differences between cells and beads  in this channel, using BD™ CompBeads can result in spillover errors for V500 and AmCyan reagents.   Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended.   Different V500 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.

560774 Rev. 2
Format Details
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V500
BD Horizon™ V500 Dye is part of the BD Horizon™ Violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 415-nm and an emission maximum (Em Max) at 499-nm. Driven by BD innovation, V500 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 500 nm (eg, a 525/50 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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V500
Violet 405 nm
415 nm
499 nm
560774 Rev.2
Citations & References
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Development References (3)

  1. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  2. 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.
  3. 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.
560774 Rev. 2

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.