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RB744 Mouse Anti-Human CD56
RB744 Mouse Anti-Human CD56
Multiparameter flow cytometric analysis using BD OptiBuild™ RB744 Mouse Anti-Human CD56 antibody (Cat. No. 757213) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Multiparameter flow cytometric analysis using BD OptiBuild™ RB744 Mouse Anti-Human CD56 antibody (Cat. No. 757213) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
Product Details
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BD OptiBuild™
N-CAM; NCAM1; NCAM-1; Neural cell adhesion molecule 1; NKH1; MSK39
Human (Tested in Development)
Mouse BALB/c X C57BL/6 IgG1, κ
KG1a Cell Line
Flow cytometry (Qualified)
0.2 mg/ml
V NK19
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 that minimize unconjugated dye and antibody.

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  4. 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.
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. 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.
  7. 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.
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. An isotype control should be used at the same concentration as the antibody of interest.
  11. 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.
  12. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
757213 Rev. 2
Antibody Details
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MY31

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.

757213 Rev. 2
Format Details
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RB744
The BD Horizon RealBlue™ 744 (RB744) 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 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 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), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).
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RB744
Blue 488 nm
498 nm
746 nm
757213 Rev.2
Citations & References
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Development References (9)

  1. 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.
  2. Edelman GM. Cell adhesion molecules.. Science. 1983; 219(4584):450-7. (Biology). View Reference
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
View All (9) View Less
757213 Rev. 2

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