Quantification of Calpains Activites by Ion - exchange Chromatography

摘要

Calpain had been reported to be a key event in the meat tenderness during postmortem storage of carcasses. Results indicated that separating the components of Ca~(2+)-dependent proteolytic system by DEAE-Sephacel ion-exchange chromatography method was an effective method. On the basis of repeated experiments, we had found a good way of calpain crude extract prapation. To purify calpain-calpastatin system, crude extract was treated with DEAE-ion exchange chromatography and then with ion strength linear gradient elution by Nad with the concentration ranged from 0 to 500 mM and Tris-HCl. The purity of calpain were tested by SDS-PAGE gel electrophoresis that indicated the extracted calpain solution had a higher purity.