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Methods Mol Biol. 2013;977:339-51. doi: 10.1007/978-1-62703-284-1_27.

Fluorescence anisotropy microplate assay to investigate the interaction of full-length steroid receptor coactivator-1a with steroid receptors.

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1
Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL, USA.

Abstract

Estrogens, acting via estrogen receptor (ER) play key roles in growth, differentiation, and gene regulation in the reproductive, central nervous, and skeletal systems. ER-mediated gene transcription contributes to the development and spread of breast, uterine, and liver cancer. Steroid receptor coactivator-1a (SRC1a) belongs to the P160 family of coactivators, which is the best known of the many coactivators implicated in ER-mediated transactivation. Binding of full-length P160 coactivators to steroid receptors has been difficult to investigate in vitro. This chapter details how to investigate the interaction of SRC1a with ER using the fluorescence anisotropy/polarization microplate assay (FAMA).

PMID:
23436375
PMCID:
PMC3708604
DOI:
10.1007/978-1-62703-284-1_27
[Indexed for MEDLINE]
Free PMC Article
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