Quantitative, solution-phase profiling of multiple transcription factors in parallel

Betul Bilgin, Li Liu, Christina Chan, S. Patrick Walton

Research output: Contribution to journalArticle

  • 7 Citations

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 proof-of-principle analyses of purified human transcription factors and breast cancer nuclear extracts. Our assay can successfully quantify transcription factors in parallel with ∼10-fold better sensitivity than current techniques. Sensitivity of the assay can be further increased by 200-fold through the use of PCR for signal amplification.

LanguageEnglish (US)
Pages2461-2468
Number of pages8
JournalAnalytical and Bioanalytical Chemistry
Volume405
Issue number8
DOIs
StatePublished - Mar 2013

Profile

Transcription Factors
Assays
Genes
Amplification
Breast Neoplasms
Polymerase Chain Reaction
DNA
Proteins

Keywords

  • Breast cancer
  • DNA binding activity
  • Magnetic beads separation
  • Parallel
  • Transcription factors

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Quantitative, solution-phase profiling of multiple transcription factors in parallel. / Bilgin, Betul; Liu, Li; Chan, Christina; Patrick Walton, S.

In: Analytical and Bioanalytical Chemistry, Vol. 405, No. 8, 03.2013, p. 2461-2468.

Research output: Contribution to journalArticle

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