Active Matrix Metalloproteinase 9 (mmP9) (APA553Hu01)

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, 92kDa 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 10pg of MMP-2 can already be detected. Briefly, various concentrations of recombinant human MMP9 (1000ng, 500ng, 100ng, 10ng) were denatured by SDS loading buffer, electrophoresed through sodium dodecylsulphate- polyacrylamide gel (SDS-PAGE; 10% gels) containing gelatin (1mg/mL) with nonreducing conditions. After renaturation, incubation and CCB-stained, active MMP2 would hydrolyze gelatin nearby, which was indicated by the white binds on the gel. In this experiment we use heat-denatured MMP9 protein as negative control, and blood sample as positive control. Result: Gelatin hydrolysis by recombinant human MMP9 (10-70kd) was shown in figure 1.

Figure. Gene Sequencing (Extract)

Figure. SDS-PAGE