Authors: Betul Bilgin Li Liu Christina Chan S Patrick Walton
Publish Date: 2013/01/30
Volume: 405, Issue: 8, Pages: 2461-2468
Abstract
Transcription factors are regulatory proteins that bind to specific sites of chromosomal DNA to enact responses to intracellular and extracellular stimuli Transcription factor signalling networks are branched and interconnected so that any single transcription factor can activate many different genes and one gene can be activated by a combination of different transcription factors Thus trying to characterize a cellular response to a stimulus by measuring the level of only one transcription factor potentially ignores important simultaneous events that contribute to the response Hence parallel measurements of transcription factors are necessary to capture the breadth of valuable information about cellular responses that would not be obtained by measuring only a single transcription factor We have sought to develop a new scalable flexible and sensitive approach to analysis of transcription factor levels that complements existing parallel approaches Here we describe proofofprinciple analyses of purified human transcription factors and breast cancer nuclear extracts Our assay can successfully quantify transcription factors in parallel with ∼10fold better sensitivity than current techniques Sensitivity of the assay can be further increased by 200fold through the use of PCR for signal amplificationWe thank all the members of the Cellular and Biomolecular Laboratory http//wwwegrmsuedu/cbl/ for their advice and support Financial support for this work was provided in part by Michigan State University the National Science Foundation CBET 0941055 the National Institutes of Health GM079688 RR024439 GM089866 DK081768 DK088251 the Michigan Universities Commercialization Initiative MUCI and the Center for Systems Biology
Keywords: