SDS-PAGE (NBC1-18526)

1. Prepare a recombinant PAI-1 with various concentrations (2,000 nM, 1,000 nM, 500 nM, 250 nM, 125 nM) in assay buffer. (Assay buffer: 50 mM Tris-HCl pH 8.0, 0.01% Tween 20)

2. Mix equal volumes of the PAI-1 protein and diluted 2 ug/ml uPA

3. Incubate at room temperature for 15 minutes.

4. After dilute incubated reaction mixtures (PAI-1, uPA) by a 5 fold dilution in assay buffer, mix 50ul of 200uM substrate (Z-GGR-AMC) and diluted mixtures.

4. Read excitation wavelength 355nm and emission wavelength 460nm for 10 minutes.