Assay of matrix metalloproteinases in substrate impregnated gels in multiwells.

ABSTRACT

A zymographic method for the assay of matrix metalloproteinases in substrate impregnated gels in multiwells has been developed for the analysis of a large number of samples at a time. Enzyme was copolymerized with 300 microliters of 10% acrylamide impregnated with gelatin substrate and incubated for 16 hr. The gels were stained with coomassie blue, destained with water and the intensity measured in a densitometer. This method was tested with pure bacterial collagenase and three different gelatinases purified from rat mammary gland. The characteristics of these enzymes such as cation dependence, inhibition and concentration dependence have been examined by this method.