BD Horizon™ RB780 Mouse Anti-Human CD337 (NKp30)
Clone p30-15 (RUO)
Multiparameter analysis of CD337 (NKp30) expression on Human peripheral blood lymphocytes. Human whole blood was stained with APC Mouse Anti-Human CD56 antibody (Cat. No. 555518) and with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plots) or BD Horizon™ RB780 Mouse Anti-Human CD337 (NKp30) antibody (Cat. No. 569205/569206; Right Plots). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plots showing the correlated expression of CD337 (NKp30) [or Ig Isotype control staining] versus CD56 (Top Plots) or side light-scatter (SSC-A) signals (Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of either intact lymphocytes or leucocyte populations, respectively. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
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Multiparameter analysis of CD337 (NKp30) expression on Human peripheral blood lymphocytes. Human whole blood was stained with APC Mouse Anti-Human CD56 antibody (Cat. No. 555518) and with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plots) or BD Horizon™ RB780 Mouse Anti-Human CD337 (NKp30) antibody (Cat. No. 569205/569206; Right Plots). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plots showing the correlated expression of CD337 (NKp30) [or Ig Isotype control staining] versus CD56 (Top Plots) or side light-scatter (SSC-A) signals (Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of either intact lymphocytes or leucocyte populations, respectively. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
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.
- 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.
- 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).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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.
- 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 U.S. patents that may apply, see bd.com/patents.
Companion Products
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The p30-15 monoclonal antibody specifically binds to CD337, also known as NKp30, a receptor found on the surface of natural killer (NK) cells. NK cells are large lymphoid cells discovered because of their ability to recognize and kill abnormal cells such as tumor and virally infected cells. NK cell immune responses are regulated by a balance of activating and inhibitory signals generated by cell surface receptors. Inhibitory receptors recognize MHC class I molecules on normal cells producing a negative signal to the NK cell. Loss of MHC class I expression in infected or transformed cells results in the loss of this negative signal leading to NK cell activation. In concert with the loss of inhibitory signals, activation signals via NK receptors such as NKp30, NKp44, NKp46, NKG2D, and NKp80 mediate the activation of NK cells. NKp30 cooperates with NKp46 and/or NKp44 in the induction of NK cell-mediated cytotoxicity against the majority of target cells.
Development References (5)
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Augugliaro R, Parolini S, Castriconi , et al. Selective cross-talk among natural cytotoxicity receptors in human natural killer cells. Eur J Immunol. 2003; 33(5):1235-1241. (Biology). View Reference
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Byrd A, Hoffmann SC, Jarahian M, Momburg F, Watzl C. Expression analysis of the ligands for the Natural Killer cell receptors NKp30 and NKp44. PLoS ONE. 2007; 2(12):e1339. (Immunogen: Blocking, ELISA, Flow cytometry). View Reference
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Flaig RM, Stark S, Watzl C. Cutting edge: NTB-A activates NK cells via homophilic interaction. J Immunol. 2004; 172(11):6524-6527. (Biology). View Reference
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Pende D, Parolini S, Pessino A, et al. Identification and molecular characterization of NKp30, a novel triggering receptor involved in natural cytotoxicity mediated by human natural killer cells. J Exp Med. 1999; 190(10):1505-1516. (Biology). View Reference
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Stark S, Flaig RM, Sandusky M, Watzl C. The use of trimeric isoleucine-zipper fusion proteins to study surface-receptor-ligand interactions in natural killer cells. J Immunol Methods. 2004; 296(1-2):149-158. (Biology). View Reference
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