The MR9-4 monoclonal antibody specifically recognizes the Vβ 5.1 and Vβ 5.2 T-cell Receptors of strains having the b haplotype (e.g., C57BL) of the Tcrb gene complex. These gene loci are deleted in mice having the a (e.g., C57BR, C57L, SJL, SWR) or c (e.g., RIII) Tcrb haplotype. Vβ5.1 and 5.2 TCR-bearing T lymphocytes are clonally eliminated, either completely or partially, in mice expressing I-E and superantigens encoded by the Mtv-1 (Mls-4a, Mlsc), Mtv-3 (Mlsc), Mtv-8 (Mlsf), Mtv-9 (Etc-1, Mlsf), Mtv-11 (Mlsf), Mtv-13 (Mls-2a, Mlsc), Mtv-27, Mtv44 , and/or Mtv-MAI endogenous provirus (e.g., A, AKR, BALB/c, C3H/He, C58, CBA/Ca, CBA/J, DBA/2, NZB, NZW). Activation of Vβ5 TCR-expressing T cells by this determinant is dependent upon presentation by I-E. Plate-bound MR9-4 antibody activates Vβ5.1 or 5.2 TCR-bearing T cells.
The antibody was conjugated to an oligonucleotide that contains an antibody clone-specific barcode (ABC) flanked by a poly-A tail on the 3' end and a PCR handle (PCR primer binding site) on the 5' end. The ABC for this antibody was designed to be used with other BD AbSeq oligonucleotides conjugated to other antibodies. All AbSeq ABC sequences were selected in silico to be unique from human and mouse genomes, have low predicted secondary structure, and have high Hamming distance within the BD AbSeq portfolio, to allow for sequencing error correction and unique mapping. The poly-A tail of the oligonucleotide allows the ABC to be captured by the BD Rhapsody™ system. The 5' PCR handle allows for efficient sequencing library generation for Illumina sequencing platforms.
NOTE: The BD Rhapsody Single-Cell Analysis System must be used with the BD Rhapsody Express Instrument.