A New Analytical Scale DNA Affinity Binding Assay for Analyses of Specific Protein–DNA Interactions
✍ Scribed by N. Vinay Kumar; Lori R. Bernstein
- Publisher
- Elsevier Science
- Year
- 2001
- Tongue
- English
- Weight
- 160 KB
- Volume
- 299
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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.