I am doing EMSA (using pierce Lightshift kit 20148) to study activation of transcription factors (NF-kB and AP-1). Every time I see the shift but the bands are always near the well and I am unable to see the supershift with the addition of antibodies against transcription factor subunits. Firstly I thought that amount of protein is too much but decresing protein changed nothing. I am using BioRad Mini protein 3 Cell electrophoresis system. Could you please suggest something?
Thanks in advance.
Problem in EMSA