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Multiparameter flow cytometric analysis using BD OptiBuild™ RB705 Mouse Anti-Human CD56 antibody (Cat. No. 757214) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
BD OptiBuild™ RB705 Mouse Anti-Human CD56
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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- 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.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- 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).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Clone MY31 specifically recognizes the human form of the 220/135 kDa heavily glycosylated antigen, CD56, found on a subpopulation of peripheral blood large granular lymphocytes which demonstrate natural killer cell activity, but not on myeloid cells, erythrocytes or B cells. This clone also cross-reacts with a subset of peripheral blood lymphocytes of baboon, and both rhesus and cynomolgus macaque monkeys. The distribution on lymphocytes is similar to that observed with peripheral blood lymphocytes from normal human donors, with a subset of CD16+ cells co-expressing CD56. In contrast to what is observed with human peripheral blood cells, however, clone MY31 also reacts with a major subset of non-human primate CD14+ monocytes. Studies in rhesus macaque monkeys suggest that CD56 reacts with monocytes and not natural killer cells.
Development References (9)
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Schubert J, Lanier LL, Schmidt RE. Cluster report: CD56. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:699-702.
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Edelman GM. Cell adhesion molecules.. Science. 1983; 219(4584):450-7. (Biology). View Reference
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Hercend T, Griffin JD, Bensussan A, et al. Generation of monoclonal antibodies to a human natural killer clone. Characterization of two natural killer-associated antigens, NKH1A and NKH2, expressed on subsets of large granular lymphocytes.. J Clin Invest. 1985; 75(3):932-43. (Biology). View Reference
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Lanier LL, Chang C, Azuma M, Ruitenberg JJ, Hemperly JJ, Phillips JH. Molecular and functional analysis of human natural killer cell-associated neural cell adhesion molecule (N-CAM/CD56). J Immunol. 1991; 146(12):4421-4426. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting, Immunoprecipitation). View Reference
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Lanier LL, Le AM, Civin CI, Loken MR, Phillips JH. The relationship of CD16 (Leu-11) and Leu-19 (NKH-1) antigen expression on human peripheral blood NK cells and cytotoxic T lymphocytes. J Immunol. 1986; 136(12):4480-4486. (Immunogen: Flow cytometry). View Reference
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Lanier LL, Testi R, Bindl J, Phillips JH. Identity of Leu-19 (CD56) leukocyte differentiation antigen and neural cell adhesion molecule. J Exp Med. 1989; 169(6):2233-2238. (Clone-specific: Immunoprecipitation). View Reference
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Phillips JH, Lanier LL. Dissection of the lymphokine-activated killer phenomenon: relative contribution of peripheral blood natural killer cells and T lymphocytes to cytolysis. J Exp Med. 1986; 164(3):814-825. (Clone-specific: Flow cytometry). View Reference
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Van Der Windt DJ, Smetanka C, Macedo C, et al. Investigation of lymphocyte depletion and repopulation using alemtuzumab (Campath-1H) in cynomolgus monkeys.. Am J Transplant. 2010; 10(4):773-783. (Clone-specific: Flow cytometry). View Reference
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Wils EJ, Aerts-Kaya FS, Rombouts EJ, et al. Keratinocyte growth factor and stem cell factor to improve thymopoiesis after autologous CD34+ cell transplantation in rhesus macaques.. Biol Blood Marrow Transplant. 2012; 18(1):55-65. (Clone-specific: Flow cytometry). View Reference
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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.