Resolution of rabbit liver cytochrome P450 3b and 3c (P450 IIC3 and IIIA4) from microsomal membrane by two‐dimensional gel electrophoresis

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Abstract
Two‐dimensional gel electrophoresis was applied to the separation of microsomal proteins and P450 isozymes. The experimental conditions of the first‐dimensional isoelectric focusing were optimized. The best results were obtained under the following conditions: (1) a mixture of two detergents (Tergitol NP 10 and CHAPS) in the focusing gel and sample buffer, (ii) a carrier ampholyte gradient between pH 6.5 and 9.0, (iii) sample loading at the anodic end of the gel, and (iv) low protein loading in the sample (below 50 μg). Four P450 forms, including P450 3b and 3c, could be resolved under these conditions.