FIG. 1.
Protease cleavage of RelSeq NH 1-385. Reaction mixtures (20 μl) containing 4 μg of purified protein, 4 mM MgCl2, 5% glycerol, 50 mM NaCl, 20 mM Tris (pH 7.9), and 1 mM EDTA were incubated at room temperature for 30 min. Subtilysin, thermolysin, trypsin, or chymotrypsin was present at the concentrations (in nanograms) indicated above the lanes. The reaction was stopped by the addition of an equal volume of 2× SDS loading buffer and boiling for 2 min, and the products were run on a 12% gel. Lane O, uncleaved protein; lane STP, protein size standards.