HepG2 cells were incubated at 37&degC for 24h with vehicle control (0 &microM) and 100 µM of fluconazole (ab141065). Increased expression of cytochrome P450 1A1 (ab3568) correlates with an increase in fluconazole concentration, as described in literature.Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10&microg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab3568 at 1/500 dilution and ab8227 at 1 &microg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

HepG2 cells were incubated at 37°C for 24h with vehicle control (0 µM) and 100 µM of fluconazole (ab141065). Increased expression of cytochrome P450 1A1 (ab3568) correlates with an increase in fluconazole concentration, as described in literature.Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab3568 at 1/500 dilution and ab8227 at 1 µg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.


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