Fig. 6.

Covalent binding of SCH 66712 to CYP2D6. Labeled SCH 66712 was incubated with Supersomes (100 pmol/ml) for 10 min at 37°C in the presence or absence of 1 mM NADPH. Samples were chilled on ice after the incubation. For SDS-PAGE, 20-μl aliquots were removed, mixed with 20 μl of gel loading buffer, and boiled for 3 min before loading on the 10% acrylamide gel. For analysis, 2 pmol of 2D6 protein were loaded on the gel. Protein was transferred from gel to nitrocellulose paper for Western blotting. A, Western blot with anti-CYP2D6 antibodies. Chemiluminescence was used for detection. B, audioradiogram of gel in A.