A nonradioactive electrophoretic mobility shift assay for measurement of pregnane X receptor binding activity to CYP3A4 response element |
| |
| Authors: | Aneta Vavrova Radim Vrzal Zdenek Dvorak |
| |
| Affiliation: | Department of Cell Biology and Genetic, Faculty of Science, Palacky University Olomouc, Slechtitelu, , Olomouc, Czech Republic |
| |
| 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 and many 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. |
| |
| Keywords: | Cytochrome P450 Drug metabolism Electrophoretic mobility shift assay Pregnane X receptor Response elements |
|
|
