Recombinant Anti-Cytochrome P450 3A4/CYP3A4 antibody [EPR6202] (ab124921)

This antibody fails to detect BOI in normal HepG2 cells which is positive reported by PMID 26089940 and 25141173

ab124921 (purified) at 1/20 dilution (1ug) immunoprecipitating Cytochrome P450 3A4/CYP3A4 in Human liver lysates.
Lane 1: Human liver lysates 10ug
Lane 2 (+): ab124921 & Human liver lysates
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124921 in Human liver lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.

HepG2 cells were incubated at 37°C for 48h with vehicle control (0 μM) and different concentrations of nicardipine hydrochloride (ab120531) in DMSO. Increased expression of Cytochrome P450 3A4/CYP3A4 (ab124921, unpurified) correlates with an increase in nicardipine hydrochloride 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 5% BSA before being incubated with ab124921 (unpurified) at 1/10000 dilution and ab9484 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.

ab124921 (unpurified), at a 1/100 dilution, staining Cytochrome P450 3A4/CYP3A4 in paraffin embedded Human liver tissue by Immunohistochemistry.

Cytochrome P450 3A4/CYP3A4 antibody ab124821 (unpurified) was used with 3D Cell Culture Clearing Kit ab243299 to penetrate, stain and clear a 3D hepatocyte cell culture.

Blue: DAPI, Red: P450

Learn more about 3D cell culture and tissue clearing kits, reagents, and protocols designed to make it easier to stain 3D cell cultures and thick tissue sections and get more data from each valuable tissue section.