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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- 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 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.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The C8 monoclonal antibody specifically recognizes C57BL/6 mouse CD161 antigen-like family member F (CD161f ). CD161f is also known as Natural killer cell surface protein NKR-P1F (Nkrp1f), and is encoded by Klrb1f (Killer cell lectin-like receptor subfamily B member 1F). CD161f is expressed on natural killer (NK) cells, and some dendritic cells and activated lymphocytes. CD161f binds to C-type lectin related protein c, d, and g (Clr-c, d, g) which are expressed on several cell types including dendritic cells, B cells, and activated T cells. CD161f-mediated signaling can reportedly costimulate T cell proliferation and IL-2 production, and may play a role in dendritic cell maturation.
Development References (9)
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Aust JG1, Gays F, Mickiewicz KM, Buchanan E, Brooks CG.. The expression and function of the NKRP1 receptor family in C57BL/6 mice.. J Immunol. 2009; 183(1):106-116. (Immunogen: Flow cytometry). View Reference
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Carlyle JR, Mesci A, Ljutic B, et al. Molecular and genetic basis for strain-dependent NK1.1 alloreactivity of mouse NK cells.. J Immunol. 2006; 176(12):7511-24. (Biology). View Reference
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Chai L, Wu S, Liu G, Wang Z, Tian W, Ma Y. OCILRP2 signaling synergizes with LPS to induce the maturation and differentiation of murine dendritic cells.. Biochem Biophys Res Commun. 2014; 446(4):836-42. (Biology). View Reference
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Iizuka K, Naidenko OV, Plougastel BF, Fremont DH, Yokoyama WM. Genetically linked C-type lectin-related ligands for the NKRP1 family of natural killer cell receptors.. Nat Immunol. 2003; 4(8):801-7. (Biology). View Reference
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Kirkham CL, Carlyle JR. Complexity and Diversity of the NKR-P1:Clr (Klrb1:Clec2) Recognition Systems. Front Biosci. 2014; 5(5):1-16. (Biology). View Reference
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Plougastel B, Matsumoto K, Dubbelde C, Yokoyama WM. Analysis of a 1-Mb BAC contig overlapping the mouse Nkrp1 cluster of genes: cloning of three new Nkrp1 members, Nkrp1d, Nkrp1e, and Nkrp1f.. Immunogenetics. 2001; 53(7):592-8. (Biology). View Reference
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Rozbeský D, Ivanova L, Hernychová L, Grobárová V, Novák P, Černý J. Nkrp1 family, from lectins to protein interacting molecules.. Molecules. 2015; 20(2):3463-78. (Clone-specific). View Reference
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Tian W, Feng B, Liou HC. Silencing OCILRP2 leads to intrinsic defects in T cells in response to antigenic stimulation.. Cell Immunol. 2005; 235(1):72-84. (Biology). View Reference
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Tian W, Nunez R, Cheng S, et al. C-type lectin OCILRP2/Clr-g and its ligand NKRP1f costimulate T cell proliferation and IL-2 production.. Cell Immunol. 2005; 234(1):39-53. (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.