Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) 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).
Product Notices
- This antibody was developed for use in flow cytometry.
- 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.
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.
The antibody was conjugated to BD Horizon™ BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (11)
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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
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.
