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. Author manuscript; available in PMC: 2009 Aug 1.
Published in final edited form as: Drug Metab Dispos. 2008 May 7;36(8):1637–1649. doi: 10.1124/dmd.107.019562

Figure 11.

Figure 11

SDS-PAGE/PhosphorImager analysis of [14C]3-OHCBZ covalent binding to mouse liver microsomal protein. Liver microsomes from Dex-treated mice were incubated as described (Materials and Methods) with freshly prepared GSH (0, 1 or 4 mM). A 50 μl-aliquot of the incubation mixture (≈ 100 pmol P450) was removed at 30 min, mixed with the SDS-PAGE loading buffer, heated at 95°C for 5 min and subjected to SDS-PAGE analyses. The gel was fixed, the signal amplified with Amersham Amplify™ fluorographic reagent, and then subjected to fluorography by PhosphorImager analyses. Under these conditions, covalent binding of [14C]3-OHCBZ to mouse liver microsomes in incubations containing NADPH was 5.32 ± 0.19 (in the absence of GSH), 0.91 ± 0.23 (with 1 mM GSH), and 0.38 ± 0.10 (with 4 mM GSH) pmol/pmol P450/30 min, respectively, and appeared to be concentrated in a single band with a molecular weight of ~55 kDa. An image of the full gel is included as supplementary information.