The UPSTATE Non-Radioactive Universal EZ-TFA Transcription Factor Assay provides a fast, sensitive method to detect specific transcription factor DNA binding activity in whole cell or nuclear extracts. This assay combines the principle of the electrophoretic mobility shift assay (EMSA) with the 96-well based enzyme-linked immunosorbent assay (ELISA), enabling manual or high-throughput sample analysis with greater sensitivity than conventional EMSA assays. After certain parameters are optimized, the entire assay takes less than 4 hours to complete with minimal hands-on time. The versatile set up allows for a flexible assay design and because the binding reaction occurs in solution, various parameters can be optimized such as probe titration, competitive oligonucleotide concentration, or treatment conditions. This kit contains the reagents and instructions necessary to develop an assay to monitor the DNA binding activity of any transcription factor of interest, or as a kit component for Upstate developed target specific Transcription Factor Assays. The Universal kit also allows for the design of native promoter capture probe sequences to interrogate specific transcription factor binding.
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