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 MR5-2 antibody reacts with the Vβ 8.1 and Vβ 8.2 T-cell Receptors (TCR), but not the Vβ 8.3 TCR, of mice having the b haplotype (e.g., A, AKR, BALB/c, CBA/Ca, CBA/J, C3H/He, C57BL, C58, DBA/1, DBA/2) of the Tcrb gene complex. The Tcrb-V8 subfamily gene loci are deleted in mice having the a (e.g., C57BR, C57L, SJL, SWR) or c (e.g., RIII) haplotype. Vβ 8.1 TCR-bearing T lymphocytes are clonally eliminated in mice expressing superantigen encoded by the Mtv-7 (Mls-1a, Mlsa), provirus (e.g., AKR, CBA/J, C58, DBA/2), and activation or elimination of Vβ 8.1 TCR-expressing T cells by this determinant is partially dependent upon presentation by I-E. Mtv-43 (e.g., MA/MyJ), Mtv-44 (e.g., NZW), and/or exogenous MMTV-SW superantigens also cause incomplete elimination of Vβ 8.1 TCR-bearing T cells. In addition to expression on conventional T lymphocytes, Vβ 8.2 is the predominant β chain of the TCR on NK-T cells. Staphylococcal enterotoxin B, in association with antigen presenting cells expressing I-A and/or I-E, stimulates lymphocytes bearing Vβ 8 TCR and selectively eliminates those T cells in vivo. Plate-bound MR5-2 antibody activates Vβ 8.1 or 8.2 TCR-bearing T lymphocytes.
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Hodes RJ, Abe R. Mouse endogenous superantigens: Ms and Mls-like determinants encoded by mouse retroviruses.. Curr Protoc Immunol. 2001; Appendix 1:Appendix 1F. (Biology). View Reference
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Hugo P, Kappler JW, Godfrey DI, Marrack PC. Thymic epithelial cell lines that mediate positive selection can also induce thymocyte clonal deletion. J Immunol. 1994; 52(3):1022-1031. (Biology). View Reference
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Kanagawa O. Antibody-mediated activation of T cell clones as a method for screening hybridomas producing antibodies to the T cell receptor. J Immunol Methods. 1988; 110(2):169-178. (Immunogen). View Reference
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Kruisbeek AM, Shevach EM. Proliferative assays for T cell function. Curr Protoc Immunol. 2004; 3:3.12.1-3.12.14. (Biology). View Reference
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Shinohara K, Ikarashi Y, Maruoka H, et al. Functional and phenotypical characteristics of hepatic NK-like T cells in NK1.1-positive and -negative mouse strains. Eur J Immunol. 1999; 29(6):1871-1878. (Biology). View Reference
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Tomonari K, Fairchild S. Positive and negative selection of Tcrb-V6+ T cells. Immunogenetics. 1992; 36(4):230-237. (Biology). View Reference
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White J, Herman A, Pullen AM, Kubo R, Kappler JW, Marrack P. The V beta-specific superantigen staphylococcal enterotoxin B: stimulation of mature T cells and clonal deletion in neonatal mice. Cell. 1989; 56(1):27-35. (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.