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RB705 Mouse Anti-Human CD8b
RB705 Mouse Anti-Human CD8b
Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Mouse Anti-Human CD8b antibody (Cat. No. 756931) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Mouse Anti-Human CD8b antibody (Cat. No. 756931) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Product Details
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BD OptiBuild™
CD8B; CD8B1; CD8 beta; CD8β; Leu2; Ly3; LYT3
Human (Tested in Development)
Mouse BALB/c IgG2a, κ
CD8+ suppressor T-cell
Flow cytometry (Qualified)
0.2 mg/ml
II T95
926
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
  7. 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.
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  12. For U.S. patents that may apply, see bd.com/patents.
756931 Rev. 2
Antibody Details
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2ST8.5H7

The 2ST8.5H7 monoclonal antibody specifically recognizes an epitope formed by the combination of CD8 alpha and beta chains. The majority of peripheral blood CD8+ T lymphocytes expresses a CD8αβ heterodimer (32, 30 kilodaltons (kDa)), while CD8+CD16+ natural killer (NK) cells and CD8+ TCR γδ+ T lymphocytes express CD8αα homodimers. The 2ST8.5H7 antibody can therefore be used to selectively bind to CD8+ T cells while excluding CD8+ NK cells. CD8 binds to class I major histocompatibility (MHC) molecules, resulting in increased adhesion between the CD8+ T lymphocytes and target cells. Binding of CD8 to class I MHC molecules enhances the activation of resting T lymphocytes. CD8 is coupled to a protein tyrosine kinase, p56lck. The CD8:p56lck complex can play a role in T-lymphocyte activation through mediation of the interactions between CD8 and the CD3/TCR complex. The CD8β antigen is present on the human suppressor/cytotoxic T-lymphocyte subset. The CD8 antigen is expressed on 19% to 48% of normal peripheral blood lymphocytes and 60% to 85% of normal thymocytes. The 2ST8.5H7 antibody crossreacts with lymphocytes of some nonhuman primate species.

756931 Rev. 2
Format Details
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RB705
The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
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RB705
Blue 488 nm
498 nm
707 nm
756931 Rev.2
Citations & References
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Development References (9)

  1. Fry TJ, Moniuszko M, Creekmore S, et al. IL-7 therapy dramatically alters peripheral T-cell homeostasis in normal and SIV-infected nonhuman primates. Blood. 2003; 101(6):2294-2299. (Clone-specific). View Reference
  2. Hambor JE, Weber MC, Tykocinski ML, Kaplan DR. Regulation of allogeneic responses by expression of CD8 alpha chain on stimulator cells.. Int Immunol. 1990; 2(9):879-83. (Clone-specific: Flow cytometry). View Reference
  3. Hori T, Cupp J, Wrighton N, Lee F, Spits H. Identification of a novel human thymocyte subset with a phenotype of CD3- CD4+ CD8 alpha + beta-1. Possible progeny of the CD3- CD4- CD8- subset.. J Immunol. 1991; 146(12):4078-84. (Clone-specific: Flow cytometry). View Reference
  4. Knowles RW. Reinherz EL, Haynes BF, Nadler LM, Bernstein ID, ed. Leukocyte Typing II. Human T Lymphocytes. New York, NY: Springer-Verlag; 1986:259-288.
  5. 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. (Biology). View Reference
  6. Moebius U. Cluster report: CD8. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:342-343.
  7. Pitcher CJ, Hagen SI, Walker JM, et al. Development and homeostasis of T cell memory in rhesus macaque. J Immunol. 2002; 168(1):29-43. (Clone-specific: Flow cytometry). View Reference
  8. 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
  9. Terry LA, DiSanto JP, Small TN, Flomenberg N. Differential expression of the CD8 and Lyt-3 antigens on a subset of human T-cell receptor γ/δ-bearing lymphocytes. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:345-346.
View All (9) View Less
756931 Rev. 2

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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.