What should I do if non-specific binding occurs in the immunoprecipitation experiment?
Use a more stringent wash buffer for washing. Add a non-ionic detergent (Tween-20 or Triton X-100) at a concentration between 0.01-0.1 to the wash buffer. If the beads are blocked before precipitation, add the same blocker to the wash buffer. Increase the number of wash steps or extend the duration of the wash steps. Reduce incubation time (beads and sample). Lower the antibody concentration. Employ a pre-clearing step to remove molecules that nonspecifically bind to protein A/protein G or to the beads themselves.
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