[26] Treatment of trypsin with TPCK

摘要: Publisher Summary This chapter discusses the procedure of treatment trypsin with 6- (1-tosylamido-2-phenyl) ethyl chloromethyl ketone (TPCK). TPCK is designed to react specifically active center chymotrypsin. completely inhibits proteolytic activity chymotrypsin but had little effect on trypsin. The a modification which involves smaller volumes reagents. 2g crystalline dissolved in 620 ml 0.001 M CaC1 2 , and pH adjusted 7.0 N NaOH. A solution added dropwise gentle stirring at room temperature. mixture stirred for total 5.5 hours periodic adjustment 7.0. then brought down 3.0 HC1, acidified dialyzed against 25 liters water (pH 3.0) 24 4°C. solvent removed by lyophilization give 1.4–1.5 g product that possesses specific benzoylarginine ester (BAEE) as high or higher than starting Trypsin preparations will readily digest heptapeptide become inactive substrate after being treated TPCK.