Soraia R Mendes, F Xavier Gomis-Rüth, Theodoros Goulas
The α2-macroglobulin (α2M) family comprises pan-peptidase inhibitors with shared structural characteristics and a common mechanism of action that play vital roles in various biological processes. This protocol describes the isolation and purification of the human homolog (hα2M) and outlines reaction conditions for evaluating its peptidase-inhibition activity in front of different substrates. It is valid for other homologues from both unicellular and multicellular organisms, providing a foundation for future research on physiological functions and potential biotechnological applications.