Prowl Lab,Rockefeller University
http://prowl.rockefeller.edu/recipes/gproteas/pronase.htm
Specificity
Pronase is a mixture of endo- and exo-proteinases. It cleaves almost any peptide bond.
pH optimum
7 - 8. Different components of the mixture may have different optima.
Stability
Pronase requires calcium ions. It retains activity in 1% SDS and 1% Triton X. Some components of the mixture are very stable to urea and guandinium HCl, but complete digestion will not occur.
Recipe
Dissolve 0.1 micromoles/microliter of substrate in 50 mM ammonium bicarbonate with 5 mM calcium chloride. Add enzyme solution to give a 100:1 ratio of substrate to enzyme. Incubate for 24 hours at 37 C. Addition of 1:20 aminopeptidase M may be necessar y to acheive complete digestion.
Garner, M.H., Garner, W.H., and Gurd, F.R.N. Recognition of primary sequence variations among sperm whale myoglobin components with successive proteolysis procedures. J. Biol. Chem. 249(1974)1513-1518.
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