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Flow cytometric analysis of CD335 (NKp46) expression on human peripheral blood lymphocytes. Whole blood was stained with BD Horizon™ BV650 Mouse Anti-Human CD335 (NKp46) antibody (Cat. No. 563230; solid line histogram) or with a BD Horizon™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; dashed line histogram). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV650 Mouse Anti-Human CD335 (NKp46)
Regulatory Status Legend
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
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.
- Brilliant Violet™ 650 is a trademark of Sirigen.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 9E2/Nkp46 monoclonal antibody specifically binds to CD335. CD335 is also known as the Natural killer cell p46-related protein (NKp46) and the Natural cytotoxicity triggering receptor 1 (NCR1). CD335 is a 46 kDa type I membrane glycoprotein that is expressed on resting and activated NK cells. Its extracellular region contains two C2-type, Ig-like domains. The transmembrane domain contains a positively charged amino acid (Arg) which could be involved in stabilizing the association with CD3ζ. Its intracellular region does not contain immunoreceptor tyrosine-based activating motifs (ITAM), but it is linked to intracytoplasmic transduction machinery by its association with CD3ζ and FcεRIγ adaptor proteins. CD335 along with NKp30 and NKp44 are referred to as natural cytotoxicity receptors (NCR). These receptors play very important roles in cells that kill virus-infected target cells, tumor cells and MHC-class I-unprotected cells.
The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. This dye is a tandem fluorochrome of BD Horizon™ BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon™ BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (4)
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Mandelboim O, Porgador A. NKp46. Int J Biochem Cell Biol. 2001; 33(12):1147-1150. (Biology). View Reference
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Nakajima H, Cella M, Bouchon A, et al. Patients with X-linked lymphoproliferative disease have a defect in 2B4 receptor-mediated NK cell cytotoxicity. Eur J Immunol. 2000; 30(11):3309-3318. (Immunogen: Flow cytometry). View Reference
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Sivori S, Pende D, Bottino C, et al. NKp46 is the major triggering receptor involved in the natural cytotoxicity of fresh or cultured human NK cells. Correlation between surface density of NKp46 and natural cytotoxicity against autologous, allogeneic or xenogeneic target cells. Eur J Immunol. 1999; 29(5):1656-1666. (Biology). View Reference
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Sivori S, Vitale M, Morelli L, et al. p46, a novel natural killer cell-specific surface molecule that mediates cell activation. J Exp Med. 1997; 186(7):1129-1136. (Biology). View Reference
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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.