The cell pellet was resuspended in 200 mL GPMV buffer containing 10 mM K-HEPES, pH 7.4, 140 mM NaCl, 10 mM KCl, and 2 mM CaCl 2. The cells were centrifuged again at 3,000 g for 10 min and then ...

Laboratory of Chemistry for Functionalized Surfaces, Division of Environmental Chemistry, Institute for Chemical Research, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan ...

C6 cells or cortical tissue were washed with ice cold PBS and lysed using lysis buffer (20 mM Tris-HCl [pH 8,0], 137 mM NaCl, 10% glycerol, 1% Triton X-100, 2 mM EDTA) with 50 µg/ml PMSF (Roche) and a ...

After washing with buffer B (40 mM Tris·HCl/150 mM NaCl/0.01% Nonidet P-40/10% glycerol) supplemented with 10 mM imidazole, His–Ub–PCNA was eluted from the bead with 250 mM imidazole in buffer B.