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Gel Shift Assay Kit

Components:        

            10x GS Buffer (low salt): 1 ml, store at 4ºC

            10x GS Buffer (high salt): 1 ml, store at 4ºC

            1 mg/ml Poly(dI-dC): 0.1 ml, store at 4ºC

            1 mg/ml Poly R-478: 0.1 ml, store at 4ºC

            1 mg/ml Salmon DNA: 0.1 ml, store at 4ºC

            GS Loading Solution: 1 ml, store at 4ºC

Product Description: Gel shift assay, also known as gel retardation assay or electrophoretic mobility shift assay (EMSA), is widely used for the detection of DNA/RNA-protein complexes. Gel shift assays performed with total or nuclear protein extracts require the use of nonspecific DNA competitors, usually polyanion polymers, to sequester undesired DNA-protein interaction. The choice of nonspecific competitors however can be tricky since all DNA-binding proteins exhibit different intrinsic binding specificities and affinities which may be inadvertently attenuated by the presence of the nonspecific competitors (Nucl. Acids Res. 20:140,1992; BioTechniques 15:390,1993). In addition, salt concentrations in the binding buffer may differentially affect DNA-protein and protein-protein interactions (Mol. Cell. Biol. 11:5090,1991). These factors, if not properly identified and controlled, may lead to irreproducible results. It is thus of benefit to use a set of dissimilar polyanion competitors and different binding conditions in the screening of crude protein extracts for novel DNA-binding activities. Our Gel Shift Assay Kit includes a set of three different polyanion polymers: poly(dI-dC), poly R-478, and sheared salmon sperm DNA. The binding assay can be performed in two different salt concentrations (10 and 100 mM KCl) to facilitate the detection of different types of higher-order DNA-protein complexes. Components of the assay kit are stable for at least one year if stored and handled properly. The product is for research use only.

Features: The Gel Shift Assay Kit allows you to use multiple nonspecific polyanion competitors under multiple binding buffer conditions for each promoter DNA/protein system. Novel or unknown DNA/RNA-binding activities associated with your favorite gene promoter can be rapidly uncovered using the optimized kit.