A new analytical scale DNA affinity binding assay for analyses of specific protein-DNA interactions.
We describe a rapid analytical assay for identification of proteins binding to specific DNA sequences. The DAPSTER assay (DNA affinity preincubation specificity test of recognition assay) is a DNA affinity chromatography-based microassay that can discriminate between specific and nonspecific protein-DNA interactions. The assay is sensitive and can detect protein-DNA interactions and larger multicomponent complexes that can be missed by other analytical methods. Here we describe in detail the optimization and utilization of the DAPSTER assay to isolate AP-1 complexes and associated proteins in multimeric complexes bound to the AP-1 DNA element.
Kumar
N V
NV
Department of Pathology and Laboratory Medicine, Texas A & M University System Health Science Center, College Station, TX 77843, USA.
Bernstein
L R
LR
D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000032:analysis*; D004268:DNA-Binding Proteins / Q000032:analysis*; D004527:Egg Proteins / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D016759:Oncogene Protein p65(gag-jun) / Q000378:metabolism; D011956:Receptors, Cell Surface; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014407:Tumor Cells, Cultured; D000072216:Zona Pellucida Glycoproteins
D004268:DNA-Binding Proteins; D004527:Egg Proteins; D008562:Membrane Glycoproteins; D016759:Oncogene Protein p65(gag-jun); D011956:Receptors, Cell Surface; D018808:Transcription Factor AP-1; D000072216:Zona Pellucida Glycoproteins; D004247:DNA