Solution: For some target proteins, background binding to beads can be decreased by increasing the NaCl or NP-40 concentration in the binding reaction or wash buffer. Problem: Immobilized bait ...

Soak the gel for 15 min at 15–25 °C in several volumes of alkaline transfer buffer (0.4 M NaOH, 1 M NaCl) with constant gentle agitation. (ii) Change the solution and continue soaking for 20 min.