Fluorescence Anisotropy Microplate Assay to Investigate the Interaction of Full-Length Steroid Receptor Coactivator-1a with Steroid Receptors

  • Chen Zhang
  • Steven K. Nordeen
  • David J. Shapiro
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 977)

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).

Key words

Estrogen receptor Steroid receptor coactivator-1a Fluorescence anisotropy Microplate LxxLL motif 

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Chen Zhang
    • 1
  • Steven K. Nordeen
    • 2
  • David J. Shapiro
    • 1
  1. 1.Department of BiochemistryUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  2. 2.University of Colorado at Denver and Health Sciences CenterAuroraUSA

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