Mmp 2 elisa kit

Following the conclusion of the experiment, the expression level of MMP-2 protein in dorsal skin tissue extracted from each HR-1 mouse was measured using MMP-2 ELISA kit (R&D System Inc., Minneapolis, MN, USA). One hundred microliters of MMP-2 coated antibody was dispensed into each microwell and incubated at 4 °C for 16 h. Each well was washed with wash buffer prior to the addition of 200 µL of assay diluent and a 1-hour incubation at room temperature. After diluting the standard solution and diluting the supernatant 20 times, the microplate was washed and 100 µL of standard and supernatant was added to each well and incubated for 2 h at room temperature. The microplate was washed, 100 µL of working detector was added to each well, and the microplate was incubated for 1 h at room temperature. After another wash, 100 µL of substrate solution was added to each well prior to incubation in a dark room for 30 min at room temperature. Finally, 50 µL of stop solution was added to each well and the absorbance was measured at 450 nm on a microplate spectrophotometer.