title: Detection of activated STAT species using electrophoretic mobility shift assay (EMSA) and potential pitfalls arising from the use of detergents.
abstract: Here we describe the preparation of nuclear extracts and the electrophoretic mobility shift assay (EMSA) for the detection of STAT species. We use the method for the investigation of STAT1 and STAT3 homo- and heterodimers and show how the preparation of the extracts can influence the distribution of the STAT species observed in the EMSA. We show that detergents can massively influence the STAT dimer distribution. Although it is unclear whether they primarily interfere with STAT DNA binding and/or whether they break up or further oligomerize STATs, the observation may also have an impact on the results of other techniques performed with detergent-containing cell lysates (e.g., coimmunoprecipitations of STATs with other proteins).
authors.affiliation: Life Sciences Research Unit, University of Luxembourg, Luxembourg, UK.
authors.forename: Serge
authors.initials: S
authors.lastname: Haan
authors.forename: Claude
authors.initials: C
authors.lastname: Haan
mesh_terms: D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D047468:Immunoprecipitation; D009841:Oligonucleotides / Q000378:metabolism; D055503:Protein Multimerization / Q000187:drug effects*; D020836:Protein Structure, Quaternary; D050791:STAT Transcription Factors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D003902:Detergents; D009841:Oligonucleotides; D050791:STAT Transcription Factors