Prowl Lab,Rockefeller University
http://prowl.rockefeller.edu/recipes/sproteas/trypsin.htm
SpecificityTrypsin cleaves very specifically at R-X and K-X bonds. If X=P, no cleavage occurs. Main trypsin preparations contain some chymotrypsin activity.
pH optimum7 - 9
StabilityTrypsin retains activity in 0.1% SDS, 1 M guanidine HCl and 30% ethanol. Trypsin is autolytic, although 20 mM calcium chloride has been reported to slow autolysis. Trypsin is irreversibly inactived at pH > 11. Trypsin is not very stable above 40 C.
RecipeDissolve the substrate in 100 mM ammonium bicarbonate at 1-10 g/l. Add the enzyme at 1:50-100 and incubate for 1-4 hours at 37 C. Much longer incubations can be used if the enzyme preparation has low chymotrypic activity.
Eggerer, J. Hysteretic behaviour of citrate synthase. Site-directed limited proteolysis. Eur. J. Biochem. 143(1984)205-212.
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