title: A nonradioactive electrophoretic mobility shift assay for measurement of pregnane X receptor binding activity to CYP3A4 response element.
abstract: The electrophoretic mobility shift assay (EMSA) is a method for the study of specific DNA–protein interactions in vitro. The pregnane X receptor (PRX) is a key xenobiotic sensor that regulates the expression of drug-metabolizing enzymes andmany other genes. Radiolabeled ³²P-DNA-probes had been used in studies of PXR-DNA interactions. There is an increasing need for nonradioactive assays, due to the health, safety and environmental issues. In the current study, we present a protocol for the nonradioactive electrophoretic mobility shift assay, allowing studying interactions between human PXR with promoter DNA sequences.
authors: Department of Cell Biology and Genetic, Faculty of Science, Palacky University Olomouc, Slechtitelu, Olomouc, Czech Republic.
authors: Aneta
authors: A
authors: Vavrova
authors: Radim
authors: R
authors: Vrzal
authors: Zdenek
authors: Z
authors: Dvorak
mesh_terms: D051544:Cytochrome P-450 CYP3A / Q000235:genetics* / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D015335:Molecular Probes / Q000235:genetics / Q000378:metabolism; D000077297:Pregnane X Receptor; D011987:Receptors, Steroid / Q000235:genetics* / Q000378:metabolism*; D020218:Response Elements
chemical_list: D015335:Molecular Probes; D000077297:Pregnane X Receptor; D011987:Receptors, Steroid; D051544:Cytochrome P-450 CYP3A