I am preparing to look at protein binding to regions of promoter DNA. I
would appreciate hearing from anyone who can suggest a systematic method
for optimizing protein binding conditions for a gel mobility shift
assay. A previous lab member here used nuclear and whole-cell extracts
and allowed 10 minutes for binding before running the labeled DNA
through a 12% acrylamide 0.5 x TBE buffered gel. Can anyone tell me if
this is even a suitable buffer for retaining protein-DNA interactions?
Thanks in advance for any assistance you can offer,
Mark Schoenbeck
Agronomy
University of Kentucky