According to the method of Gómez-Guillén (20 (link)), weighing 2 g of the gel sample, adding 10 mL of S1 solution (0.05 mol/L NaCl, 10 ml), S2 solution (0.6 mol/L NaCl), S3 solution (0.6 mol/L NaCl + 1.5 mol/L urea), Solution S4 (0.6 mol/L NaCl + 8 mol/L urea), solution S5 (0.6 mol/L NaCl + 8 mol/L urea + 0.5 mol/L β- mercaptoethanol), homogenized at low speed for 2 min, placed in an environment at 4°C for 1 h, and centrifuged at 10,000 × g for 15 min at 4°C. The supernatant was collected, and the content of protein was determined by biuret method. The chemical acting force is calculated according to formulae 1-2)–1-5:
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