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.
- 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.
Companion Products
The CD1d42 monoclonal antibody recognizes CD1d. Cell surface CD1d is structurally homologous to Class I MHC molecules. It consists of a glycosylated type I transmembrane α chain (43-49 kDa) that is non-covalently associated with β2-microglobulin. CD1d is a member of the CD1 family of molecules, which belong to the immunoglobulin superfamily. Sequence homology data classifies the CD1 molecules into two groups. Group 1 includes CD1a, CD1b and CD1c molecules; group 2 includes CD1d molecules and their homologs in other species. CD1d is expressed on cortical thymocytes, B cells, dendritic cells, monocytes, and some nonlymphoid cells including intestinal epithelial cells, hepatocytes and keratinocytes. It is not expressed on resting mature T cells. Studies suggest that CD1d participates in lipid antigen presentation to CD1d-restricted NKT cells.
Development References (6)
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Exley M, Garcia J, Wilson SB, et al. CD1d structure and regulation on human thymocytes, peripheral blood T cells, B cells and monocytes. Immunology. 2000; 100(1):37-47. (Immunogen: Flow cytometry, Immunohistochemistry, Immunoprecipitation). View Reference
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Hong S, Scherer DC, Singh N. Lipid antigen presentation in the immune system: lessons learned from CD1d knockout mice. Immunol Rev. 1999; 169:31-44. (Biology). View Reference
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Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
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Ronger-Savle S, Valladeau J, Claudy A, et al. TGFbeta inhibits CD1d expression on dendritic cells. J Invest Dermatol. 2005; 124(1):116-118. (Clone-specific: Flow cytometry). View Reference
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Somnay-Wadgaonkar K, Nusrat A, Kim HS. Immunolocalization of CD1d in human intestinal epithelial cells and identification of a beta2-microglobulin-associated form. 1999; 11(3):383-392. (Biology). View Reference
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Zola H, Swart B, Nicholson I, Voss E. CD1a–e. In: Zola H. Leukocyte and Stromal Cell Molecules : the CD Markers. Hoboken, N.J.: Wiley-Liss; 2007:42-43.
Please refer to Support Documents for Quality Certificates
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.