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 CompBead to 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
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- 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.
- 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 http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- BD Horizon Brilliant Ultraviolet 615 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.
Companion Products
The 165903 monoclonal antibody specifically recognizes UL16-binding protein 2, 5, and 6 (ULBP-2/5/6). UL16 is a human cytomegalovirus (HCMV) glycoprotein that can bind to human ULBP proteins which share homology with MHC class I proteins. ULBP-2 is also known as RAET1H (retinoic acid early transcript), NKG2DL2 (NKG2D ligand 2), and ALCAN-alpha. ULBP-5 is likewise known as RAET1G whereas ULBP-6 is also referred to as RAET1L. The ULBP-2/5/6 proteins possess only the alpha 1 and alpha 2 Ig-like domains and have no capacity to bind peptides or interact with beta 2 microglobulin. All three proteins can be expressed on the cell surface as glycophosphatidylinositol (GPI)-linked forms whereas ULBP2 and ULBP5 also exist as transmembrane-anchored forms. These proteins may be expressed on various cells types during development or cells experiencing stress caused by high proliferative activity, infection, transformation, or exposure to mediators in inflammatory or autoimmune diseases. The ULBP-2/5/6 proteins can bind to NKG2D and thereby transduce activating signals into NK cells and costimulatory signals into activated CD8+ T cells resulting in enhanced cell-mediated cytotoxicity or cytokine production. The expression of ULBP-2/5/6 proteins on infected or tumor cells can play an important role in immune surveillance and homeostasis.
The antibody was conjugated to BD Horizon BUV615 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 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP. Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).
Development References (5)
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Cosman D, Müllberg J, Sutherland CL, et al. ULBPs, novel MHC class I-related molecules, bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor.. Immunity. 2001; 14(2):123-33. (Biology). View Reference
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Lanier LL. NKG2D Receptor and Its Ligands in Host Defense.. Cancer Immunol Res. 2015; 3(6):575-82. (Biology). View Reference
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Raulet DH, Gasser S, Gowen BG, Deng W, Jung H. Regulation of ligands for the NKG2D activating receptor. Annu Rev Immunol. 2013; 31:413-441. (Biology). View Reference
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Uhlenbrock F, Hagemann-Jensen M, Kehlet S, Andresen L, Pastorekova S, Skov S. The NKG2D ligand ULBP2 is specifically regulated through an invariant chain-dependent endosomal pathway.. J Immunol. 2014; 193(4):1654-65. (Clone-specific: Flow cytometry). View Reference
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Wang R, Jaw JJ, Stutzman NC, Zou Z, Sun PD. Natural killer cell-produced IFN-γ and TNF-α induce target cell cytolysis through up-regulation of ICAM-1.. J Leukoc Biol. 2012; 91(2):299-309. (Biology). View Reference
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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.