My experiment worked when I used 32P but not now with the LightShift Kit. What is wrong?
The LightShift Kit is used to detect a biotinylated probe in an EMSA. The binding conditions for each experiment must be optimized for the proteins tested. If an experiment was working when 32P labeled probes were used, then use the same binding conditions from those previous experiments and only substitute the biotinylated probe in place of the 32P labeled probe. While commonly used reaction components are provided with the LightShift Chemiluminescent EMSA Kit, these reagents may not be applicable in all situations.
