Multiparameter flow cytometric analysis of CD56 (NCAM-1) expression on Human peripheral blood lymphocytes. Human whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD16 antibody (Cat. No. 563785) and with either BD Horizon™ RB545 Mouse IgG1, κ Isotype Control (Cat. No. 569284; Left Plots) or BD Horizon™ RB545 Mouse Anti-Human CD56 (NCAM-1) antibody (Cat. No. 569263/569264; Right Plots). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plots showing the correlated expression of CD56 (NCAM-1) [or Ig Isotype control staining] versus CD16 (Top Plots) or side light-scatter (SSC-A, Bottom Plots) signals were derived from gated events with the forward and side-light scatter characteristics of intact lymphocytes (Top Plots) or intact leucocytes (Bottom Plots). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 Software.
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Multiparameter flow cytometric analysis of CD56 (NCAM-1) expression on Human peripheral blood lymphocytes. Human whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD16 antibody (Cat. No. 563785) and with either BD Horizon™ RB545 Mouse IgG1, κ Isotype Control (Cat. No. 569284; Left Plots) or BD Horizon™ RB545 Mouse Anti-Human CD56 (NCAM-1) antibody (Cat. No. 569263/569264; Right Plots). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plots showing the correlated expression of CD56 (NCAM-1) [or Ig Isotype control staining] versus CD16 (Top Plots) or side light-scatter (SSC-A, Bottom Plots) signals were derived from gated events with the forward and side-light scatter characteristics of intact lymphocytes (Top Plots) or intact leucocytes (Bottom Plots). Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 Software.
Product Details
BD Horizon™
NCAM1; NCAM-1; Neural cell adhesion molecule 1; NCAM; NKH1; MSK39
Human (QC Testing), Rhesus,Cynomolgus,Baboon (Reported)
Mouse IgG1, κ
Human NK Cells
Flow cytometry (Routinely Tested)
5 µl/test
V NK75
4684
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® 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.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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).
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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
RB545 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
569284
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Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
BUV395 Mouse Anti-Human CD16 RUO
Size
100 Tests
Cat No.
563785
569263 Rev. 2
Antibody Details
B159
The B159 monoclonal antibody specifically binds to CD56. CD56 is a heavily glycosylated adhesion protein that is present on a subpopulation of peripheral blood large granular lymphocytes that demonstrate natural killer activity. CD56 is also expressed on a subset of T cells but is not expressed on myeloid cells, erythrocytes or B cells. This antigen is a pan-NK-cell marker. CD56 is virtually identical to an isoform of the neural cell adhesion molecule (NCAM), a structure mediating homotypic and heterotypic cell-cell interactions.
569263 Rev. 2
Format Details
RB545
The BD Horizon RealBlue™ 545 (RB545) fluorochrome is part of the BD family of blue (488-nm) laser dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 496-nm and an emission maximum (Em Max) at 545-nm. Driven by BD innovation, RB545 can be used on spectral cytometers and is designed to be excited by the blue (488-nm) laser with minimal excitation by the 561-nm yellow-green laser. RB545 has minimal spillover into yellow-green detectors and a brightness level similar to FITC. Given its unique emission max, on a spectral instrument RB545 can be used simultaneously with FITC and PE to provide an additional color off of the blue laser. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
RB545
Blue 488 nm
496 nm
545 nm
569263 Rev.2
Citations & References
View product citations for antibody "569263" on CiteAb
Development References (6)
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Bennett IM, Zatsepina O, Zamai L, Azzoni L, Mikheeva T, Perussia B. Definition of a natural killer NKR-P1A+/CD56-/CD16- functionally immature human NK cell subset that differentiates in vitro in the presence of interleukin 12. J Exp Med. 1996; 184(5):1845-1856. (Biology: Flow cytometry). View Reference
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Crotta S, Stilla A, Wack A, et al. Inhibition of natural killer cells through engagement of CD81 by the major hepatitis C virus envelope protein. J Exp Med. 2002; 195(1):35-41. (Clone-specific: Flow cytometry). View Reference
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Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
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Maraiani E, Cattini L, Piacentini A, Sgobbi S, Facchini A. Distribution of Workshop NK-cell and CD56 mAb in human peripheral blood lymphocytes during aging. In: 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; 1995:1394-1397.
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Ritz J, Trinchieri G, Lanier LL. NK-cell Antigens: Section Report. In: 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; 1995:1367-1372.
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Robertson MJ, Cochran KJ, Ritz J. Characterization of surface antigens expressed by normal and neoplastic human NK cells. In: 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; 1995:1374-1377.
569263 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.