Detection of gelatinolytic activity of ab125181 MMP2 protein. 10 ng of ab125181 in 25 µl of 25 mM Tris-glycine buffer (pH 6.8) was mixed with 2x SDS-sample buffer (without ß-mercaptoethanol) and incubated at 37°C for 10 minutes. Samples were loaded on a 10% acrylamide-Tris-glycine gel containing 0.1% gelatin and electrophoresed using Tris-glycine (pH 8.3) with 0.1% SDS. Electrophoresis was performed at 90V for 2 hours. The gel was washed 3 times in 40-50 ml of 2.5% Triton X-100 for 15 minutes. The gel was immersed in 40-50 ml of 50 mM Tris-glycine (pH 7.5), 100 mM NaCl, 4 mM CaCl 2 and 0.02% Triton X-100 and incubated at 37°C for 16 hours. For detection of the gelatinolytic activity, the gel was stained with 0.05% Commassie Blue. ab125181 produced a prominent gelatinolytic band corresponding to a molecular mass of ~ 68 kDa. See Abreview