BD OptiBuild™ BUV563 Rat Anti-Mouse Vα 8.3 T-Cell Receptor

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.

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.