The NFATc1 TransFactor Kit provides a quick and sensitive ELISA-based method to detect DNA binding by the transcription factor NFATc1, without radioactive labeling. The kit contains a 96-well plate for measuring the DNA-binding behavior of the NFATc1 transcription factor. Individual wells have been precoated with the DNA consensus binding sequence for NFATc1. 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 the primary antibody specific for NFATc1.
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.