title: Southwestern Blotting Assay.
abstract: Southwestern blotting is a technique used to study DNA-protein interactions. This method detects specific DNA-binding proteins by incubating radiolabeled DNA with a gel blot, washing, and visualizing through autoradiography. A blot resulting from 1-dimensional SDS-PAGE reveals the molecular weight of the binding proteins. To increase separation and determine isoelectric point a 2-dimensional gel can be blotted. Additional dimensions of electrophoresis, such as a gel shift (EMSA), can precede isoelectric focusing and SDS-PAGE to further improve separation. Combined with other techniques, such as mass spectrometry, the DNA-binding protein can be identified.
authors.affiliation: Department of Chemistry, University of Texas at San Antonio, One UTSA Circle, San Antonio, TX, 78249, USA.
authors.forename: Yinshan
authors.initials: Y
authors.lastname: Jia
authors.affiliation: Department of Chemistry, University of Texas at San Antonio, One UTSA Circle, San Antonio, TX, 78249, USA.
authors.forename: Linda
authors.initials: L
authors.lastname: Nagore
authors.affiliation: Department of Chemistry, University of Texas at San Antonio, One UTSA Circle, San Antonio, TX, 78249, USA. harry.jarrett@utsa.edu.
authors.forename: Harry
authors.initials: H
authors.lastname: Jarrett
mesh_terms: D026501:Blotting, Southwestern / Q000379:methods*; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D013058:Mass Spectrometry; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; D014157:Transcription Factors; D004247:DNA
keywords: DNA; Immunoblots; Nitrocellulose membrane; Oligonucleotide; PVDF membrane; Promoter; Regulation; Response elements; Southwestern blotting; Transcription