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BUV737 Mouse Anti-Human CD56
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This product is the replacement for [564447].
BUV737 Mouse Anti-Human CD56

Two color flow cytometric analysis of CD56 expression on human peripheral blood lymphocytes.  Human whole blood was stained with FITC Mouse Anti-Human CD16 antibody (Cat. No. 555406/556618/560996) and either BD Horizon™ BUV737 Mouse IgG2b, κ Isotype Control (Cat. No. 612768; Left Plot) or BD Horizon BUV737 Mouse Anti-Human CD56 antibody (Cat. No. 612766/612767; Right Plot).  The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202).  A two-color flow cytometric contour plot showing the correlated expression of CD56 (or Ig Isotype control staining) versus CD16 was derived from gated events with the forward and side light-scatter characteristic of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Two color flow cytometric analysis of CD56 expression on human peripheral blood lymphocytes.  Human whole blood was stained with FITC Mouse Anti-Human CD16 antibody (Cat. No. 555406/556618/560996) and either BD Horizon™ BUV737 Mouse IgG2b, κ Isotype Control (Cat. No. 612768; Left Plot) or BD Horizon BUV737 Mouse Anti-Human CD56 antibody (Cat. No. 612766/612767; Right Plot).  The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202).  A two-color flow cytometric contour plot showing the correlated expression of CD56 (or Ig Isotype control staining) versus CD16 was derived from gated events with the forward and side light-scatter characteristic of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Product Details
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BD Horizon™
NCAM1; NCAM-1; NCAM; Leu-19; Neural cell adhesion molecule 1; NKH1; MSK39
Human (QC Testing)
Mouse BALB/c IgG2b, κ
Immunoaffinity-enriched adult human brain NCAM
Flow cytometry (Routinely Tested)
5 µl
V NK60
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 with BD Horizon BUV737 under optimum conditions, and unconjugated antibody and free BD Horizon BUV737 were 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. This will ensure that BD CompBeads are appropriate for your specific cellular application.

     For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

     Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. BD Horizon Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
  6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
612766 Rev. 1
Antibody Details
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NCAM16.2

The NCAM16.2 monoclonal antibody specifically binds to human CD56. It recognizes an extracellular immunoglobulin-like domain common to 120, 140, and 180 kDa forms of CD56, also known as the neural cell adhesion molecule (NCAM), NKH1 or MSK39. The CD56 antigen is expressed on approximately 10% to 25% of peripheral blood lymphocytes. It is present on essentially all resting and activated CD16+ natural killer (NK) lymphocytes and approximately 5% of CD3+ peripheral blood lymphocytes. CD3+ CD56+ T lymphocytes comprise a unique subset of cytotoxic T lymphocytes that mediates non-major histocompatibility complex (MHC)-restricted cytotoxicity. CD56 antigen density on NK lymphocytes increases upon cellular activation. The CD56 antigen is involved in neuronal homotypic cell adhesion and cell differentiation during embryogenesis. CD16+ CD56+ NK cells demonstrate reciprocal transfer of an activation state with dendritic cells.

The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 737 nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 nm filter.  Due to the excitation of the acceptor dye by the red laser line, there may be significant spillover into red laser detectors with filters in the 700-720 nm range.

612766 Rev. 1
Format Details
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BUV737
The BD Horizon Brilliant™ Ultraviolet 737 (BUV737) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 735-nm. BUV737, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 740-nm (e.g., 740/35 bandpass filter). The acceptor dye can be excited by the Red (628–640nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV737
Ultraviolet 355 nm
350 nm
735 nm
612766 Rev.1
Citations & References
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Development References (14)

  1. 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). View Reference
  2. Campbell JJ, Qin S, Unutmaz D, et al. Unique subpopulations of CD56+ NK and NK-T peripheral blood lymphocytes identified by chemokine receptor expression repertoire. J Immunol. 2001; 166(11):6477-6482. (Biology). View Reference
  3. Cooper MA, Fehniger TA, Caligiuri MA. The biology of human natural killer–cell subsets. Trends Immunol. 2001; 22(11):633-640. (Biology). View Reference
  4. Cunningham BA, Hemperly JJ, Murray BA, Prediger EA, Brackenbury R, Edelman GM. Neural cell adhesion molecule: structure, immunoglobulin-like domains, cell surface modulation, and alternative RNA splicing. Science. 1987; 236(4803):799-806. (Biology). View Reference
  5. Edelman GM. Cell adhesion molecules in the regulation of animal form and tissue pattern. Annu Rev Cell Biol. 1986; 2:81-116. (Biology). View Reference
  6. Galandrini R, Tassi I, Mattia G, et al. SH2-containing inositol phosphatase (SHIP-1) transiently translocates to raft domains and modulates CD16-mediated cytotoxicity in human NK cells. Blood. 2001; 100(13):4581-4589. (Biology). View Reference
  7. Gerosa F, Baldani-Guerra B, Nisii C, Marchesini V, Carra G, Trinchieri G. Reciprocal activating interaction between natural killer cells and dendritic cells. J Exp Med. 2002; 195(3):327-333. (Biology). View Reference
  8. 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. (Immunogen: ELISA, Flow cytometry). View Reference
  9. 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. (Biology). View Reference
  10. 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. (Biology). View Reference
  11. Nitta T, Yagita H, Sato K, Okumura K. Involvement of CD56 (NKH-1/Leu-19 antigen) as an adhesion molecule in natural killer–target cell interaction. J Exp Med. 1989; 170(5):1757-1761. (Biology). View Reference
  12. 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. (Biology). View Reference
  13. 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.
  14. Schubert W, Zimmermann K, Cramer M, Starzinski-Powitz A. Lymphocyte antigen Leu-19 as a molecular marker of regeneration in human skeletal muscle. Proc Natl Acad Sci U S A. 1989; 86(1):307-311. (Biology). View Reference
View All (14) View Less
612766 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.