TransFactor kits provide a quick and sensitive ELISA-based method to detect DNA binding by a particular transcription factor, without radioactive labeling. The kits contain a 96-well plate for measuring the DNA-binding behavior of the transcription factor. Individual wells in the plate have been precoated with a consensus DNA binding sequence for the transcription factor. To perform an assay, add nuclear or cell extract from mammalian cells to the wells and incubate to allow the transcription factor to bind to its sequence. Wash away the unbound proteins, and add theprimary antibody specific for the transcription factor of interest.

The TransFactor NFATc1 Kit is available in a chemiluminescent format that uses a horseradish peroxidase-conjugated secondary antibody and chemiluminescent substrate. You can immediately measure chemiluminescent intensity using any standard luminometer or CCD camera. Increasing the amount of nuclear extract containing the transcription factor added to the well will increase the signal. By adding competitor oligo, the signal is decreased, indicating the specificity of the interaction. For quantitative results, a standard curve can be generated using purified transcription factor.