Assay of matrix metalloproteinases in substrate impregnated gels in multiwells

A zymographic method for the assay of matrix metalloproteinases in substrat e impregnated gels in multiwells has been developed for the analysis of a l arge number of samples at a time. Enzyme was copolymerised with 300 mu l of 10% acrylamide impregnated with gelatin substrate and incubated for 16 hr. The gels were stained with coomassie blue, destained with water and the in tensity measured in a densitometer. This method was tested with pure bacter ial collagenase and three different gelatinases purified fi om rat mammary gland. The characteristics of these enzymes such as cation dependence, inhi bition and concentration dependence have been examined by this method.