   7.4.2.8 | insoluble PcsN-PcsL aggregate from pellet by treatment with unfolding buffer containing 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 6 M guanidine hydrochloride for 30 min. For proper refolding, five times in volume refolding buffer, containing 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 10% glycerol, is added and left for overnight at 4°C on a stirrer rotor. Insoluble protein aggregates are removed by centrifugation and the supernatant containing soluble proteins is subjected to dialysis in dialysis buffer containing 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 10% glycerol, to remove guanidine hydrochloride |
752143 |
