Characterization of the epitope on murine T-cell receptor (TCR) alpha proteins recognized by H28-710 monoclonal antibody

Antigen recognition by alphabeta T lymphocytes is mediated via the multisubunit T-cell receptor (TCR) complex consisting of invariant CD3-gamma,delta,epsilon, and zeta chains associated with clonotypic TCRalpha,beta molecules. In the current report, we evaluated the molecular basis for recognition of murine TCRalpha proteins by H28-710 monoclonal antibody (MAb), specific for the constant region of murine TCRalpha chains. H28-710 is widely used in the study of the TCR complex as it is the only reagent currently available that recognizes all murine TCRalpha proteins, regardless of their clonotype. These data show that H28-710 is useful for the immunoprecipitation of TCRalpha proteins not associated with CD3 subunits, and that H28-710 effectively recognizes denatured TCRalpha proteins synthesized in several different cell types. Most importantly, these results demonstrate that H28 binding involves a serine/threonine-rich region between amino acids 150-177 on murine TCRalpha polypeptides.