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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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.
Companion Products
The KT50 antibody reacts with some members of the Vα 8 T-cell Receptor (TCR) subfamily of mice having the a, b, c, and d haplotypes of the Tcra gene complex (e.g., all strains tested). It recognizes an epitope in the CDR1 of Vα 8.3, but not Vα 8.2, TCR subfamily member, as does the B21.14 mAb (Cat. no. 553374). Site-directed mutagenesis has identified three amino acids which are necessary for antibody reactivity and which are unique to Vα 8.3 among the five functional Vα 8 TCR subfamily members. On a common H-2[k] background, the frequency of Vα 8.3 TCR-bearing T lymphocytes is higher in Tcra[a/a] mice than in Tcra[a/b] mice. Furthermore, studies of congenic strains suggest that CD8+ Vα 8.3 TCR-bearing T lymphocytes undergo negative selection in mice expressing MHC class I antigens of the H-2[d] haplotype.
Development References (4)
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Brodnicki TC, Holman PO, Kranz DM. Reactivity and epitope mapping of single-chain T cell receptors with monoclonal antibodies. Mol Immunol. 1996; 33(3):253-263. (Biology). View Reference
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Tomonari K, Fairchild S, Rosenwasser OA. Influence of viral superantigens on V beta- and V alpha-specific positive and negative selection. Immunol Rev. 1993; 131:131-168. (Biology). View Reference
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Tomonari K, Lovering E, Fairchild S, Spencer S. Two monoclonal antibodies specific for the T cell receptor V alpha 8. Eur J Immunol. 1989; 19(6):1131-1135. (Immunogen). View Reference
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Tomonari K. Negative selection of Tcra-V8+CD8+ T cells by MHC class I molecules. Immunogenetics. 1992; 35(5):291-295. (Biology). View Reference
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.