Quantitative, solution-phase profiling of multiple transcription factors in parallel.
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.
Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI 48824, USA.
Betul
B
Bilgin
Li
L
Liu
Christina
C
Chan
S Patrick
SP
Walton
D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D014157:Transcription Factors / Q000032:analysis* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D059467:Transcriptome
D014157:Transcription Factors