Hi All, I wanted some advice regarding mapping out Protein-peptide interaction. The peptide is a 12 mer and the protein is 15kDa. Invivo studies suggest that the peptide is binds the protein and helps in transport. Hence I feel it would perhaps transient binding. I know that I should do ITC or BIAcore to show binding, but before going to those techniques, I feel, running a native gel would perhaps help. So the native gel can have lane1: protein, lane 2: peptide, lane 3: protein+peptide.
If the protein binds to the peptide then I should not see a band corresponding to the peptide in lane 3. But before I start this experiment, I wonder if any body has run 12 mer peptide on native gel, How long should I run... How much quantity of peptide I should for the gel. I wont be able to do western or pull-down, Equipment for native gel is available to me. kindly advice, regards Anita
