Results obtained with the V79 cells were confirmed using microsomal preparation of yeast cells expressing the variants CYP2D6*1 and CYP2D6*17 and CYP2D6 inhibitor quinidine.
Taken together with the reported inhibition of CYP activities by organic solvents using microsomal fractions (Chauret et al., 1998), the mechanisms of action for this inhibition is unclear.
Furthermore, recent studies on this type of binding in various strains of mice and rats of both sexes and using microsomal preparations from various organs are briefly reviewed.
Both oxidation and reduction of glucocorticoids were detected using microsomal 11βHSD1 with reduction being the preferred direction (lower Km and higher Vmax, Table 2).
Lu et al. [ 39] recently observed inhibition of aromatase by tamNox, NDtam, and endoxifen using microsomal incubations.
The prediction of in vivo intrinsic clearance of cytochrome P450 (CYP) substrates in the rat from studies using microsomal preparations and isolated hepatocytes is elaborated in the chapter.
Protein quantification and normalization Determine the protein concentrations of microsomal extracts using the Pierce BCA Protein Assay kit (23225, Thermo Scientific Inc., Rockford, Illinois, USA) according to the manufacturer's instructions with the following modifications: Centrifuge samples and standards for 30 s, 13,000 × g.
Test 7-μL samples of microsomal extracts (in duplicate) and each standard (expect 500 μg/mL or higher for microsomal extracts).
To examine if the D. melanogaster protein is indeed posttranslationally modified with N-glycans, microsomal extracts and conditioned media of S2-dCREG cells were treated with peptide N-glycosidase F (PNGase F).
The pellet was carefully resuspended in 0.5 ml extraction buffer, and aliquots of 50 μl extracts, "microsomal extracts", were frozen and stored at -70°C.
