Can anyone help in relation to a Protease Assay? - (Nov/02/2011 )
I've to carry out a protease assay using skimmed milk agar to determine protease activity. I've also to identify the proteases produced using inhibitors. EDTA, E65 and Aprotinin. But i have no idea how to prepare these inhibitor solutions. I know i need 10mM EDTA, 100uM of Aprotinin and 1mM E64. In the past these solutions have been prepared prior to experiment by someone else, and i just injected 10ul into the well. So i've no idea what they look like and how to prepare them to use in my experiment. Please if anyone could explain asap would be a great help as i don't have any idea.
edta is soluble in water when adjusted to pH 8 (or use disodium edta (should dissolve, to ~0.1M, without adjusting pH)).
aprotinin is soluble in water.
e64 is soluble in dmso to 25mg/ml or buffer to 20mg/ml.