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Two-color flow cytometric analysis of CD337 (NKp30) expression on human peripheral blood lymphocytes. Human whole blood was stained with FITC Mouse Anti-Human CD56 antibody (Cat. No. 562794) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon™ BV421 Mouse Anti-Human CD337 (NKp30) antibody (Cat. No. 563385; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric dot plots show the correlated expression patterns of CD337 (NKp30) (or Ig Isotype control staining) versus CD56 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV421 Mouse Anti-Human CD337 (NKp30)
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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
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD Optibuild Brilliant reagents) 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).
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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.
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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For Research Use Only. Not for use in diagnostic or therapeutic procedures.