Electrophoretic Mobility Shift Assay (EMSA) and Supershift Assay of Cytochrome P450 2B6 in Response to Estrogen

Electrophoretic mobility shift assay (EMSA) is an invaluable tool to study interaction of proteins with DNA. Estrogens are major female hormones and modulate biological function through estrogen receptor (ER). ER regulates its target gene expression via the classical mechanism in which ER directly b...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1366; p. 41
Main Authors Koh, Kwi Hye, Jeong, Hyunyoung
Format Journal Article
LanguageEnglish
Published United States 2016
Subjects
Binding Sites
Binding, Competitive
Cytochrome P-450 CYP2B6 - genetics
Cytochrome P-450 CYP2B6 - metabolism
Electrophoretic Mobility Shift Assay
Estrogen Receptor alpha - agonists
Estrogen Receptor alpha - metabolism
Estrogens - pharmacology
Gene Expression Regulation, Enzymologic - drug effects
Hep G2 Cells
Hepatocytes - drug effects
Hepatocytes - enzymology
Humans
Promoter Regions, Genetic
Protein Binding
Transcription Factor AP-1 - metabolism
Workflow
Estrogen receptor
Supershift
CYP2B6
AP-1
EMSA
Estrogen
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Summary:Electrophoretic mobility shift assay (EMSA) is an invaluable tool to study interaction of proteins with DNA. Estrogens are major female hormones and modulate biological function through estrogen receptor (ER). ER regulates its target gene expression via the classical mechanism in which ER directly binds to its target gene promoter or the nonclassical mechanism involving tethering of ER to other transcription factors (such as AP-1 proteins). Here, we describe the EMSA to examine the nonclassical mechanism of ER action in regulation of a gene CYP2B6 by using competition and supershift assays.
ISSN:1940-6029
DOI:10.1007/978-1-4939-3127-9_5