Gene Regulation pp 339-351

Part of the Methods in Molecular Biology book series (MIMB, volume 977)

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

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 

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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