Mechanism:MMP9 is a zinc-dependent enzymes capable of cleaving components of the extracellular matrix, which belongs to the matrix metalloproteinase (MMP) family . It is a gelatinase A, 92 kDa type IV collagenase which can hydrolyze gelatin under certain conditions. Gelatin zymography is mainly used for the detection of the gelatinases, MMP-2 and MMP-9 and It is extremely sensitive because levels of 10 pg of MMP-2 can already be detected .Briefly，various concentrations of recombinant mouse MMP9 (100ng, 50ng, 25ng, 12.5ng, 6.25ng, 3.1ng, 1.5ng and 0.7ng) were denatured by SDS loading buffer, electrophoresed through sodium dodecylsulphate–polyacrylamide gel (SDS–PAGE; 15% gels) containing gelatin (1 mg/ml) with nonreducing conditions. After renaturation, incubation and CCB-stained, active MMP9 would hydrolyze gelatin nearby, which was indicated by the white binds on the gel. In this experiment we use trypsin as positive control.
Result: Gelatin hydrolysis by recombinant mouse MMP9(10-70kd) was shown in figure 1.