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.

    Original languageEnglish (US)
    Pages (from-to)2461-2468
    Number of pages8
    JournalAnalytical and Bioanalytical Chemistry
    Volume405
    Issue number8
    DOIs
    StatePublished - Mar 2013

    Profile

    Transcription Factors
    Genes
    Cyclic AMP Receptor Protein
    Beryllium
    Breast Neoplasms
    Polymerase Chain Reaction
    DNA
    Proteins
    Buccal Administration
    Amplification

    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

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

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

    Research output: Contribution to journalArticle

    @article{57285088745a4f4fab7b04861c2c830f,
    title = "Quantitative, solution-phase profiling of multiple transcription factors in parallel",
    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.",
    keywords = "Breast cancer, DNA binding activity, Magnetic beads separation, Parallel, Transcription factors",
    author = "Betul Bilgin and Li Liu and Christina Chan and {Patrick Walton}, S.",
    year = "2013",
    month = "3",
    doi = "10.1007/s00216-013-6712-9",
    volume = "405",
    pages = "2461--2468",
    journal = "Fresenius Zeitschrift fur Analytische Chemie",
    issn = "0016-1152",
    publisher = "Springer Verlag",
    number = "8",

    }

    TY - JOUR

    T1 - Quantitative, solution-phase profiling of multiple transcription factors in parallel

    AU - Bilgin,Betul

    AU - Liu,Li

    AU - Chan,Christina

    AU - Patrick Walton,S.

    PY - 2013/3

    Y1 - 2013/3

    N2 - 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.

    AB - 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.

    KW - Breast cancer

    KW - DNA binding activity

    KW - Magnetic beads separation

    KW - Parallel

    KW - Transcription factors

    UR - http://www.scopus.com/inward/record.url?scp=84878384378&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84878384378&partnerID=8YFLogxK

    U2 - 10.1007/s00216-013-6712-9

    DO - 10.1007/s00216-013-6712-9

    M3 - Article

    VL - 405

    SP - 2461

    EP - 2468

    JO - Fresenius Zeitschrift fur Analytische Chemie

    T2 - Fresenius Zeitschrift fur Analytische Chemie

    JF - Fresenius Zeitschrift fur Analytische Chemie

    SN - 0016-1152

    IS - 8

    ER -