FAQ: Why is the induced protein insoluble (C2529)?

T7 expression often leads to very high production of protein that can result in the target protein becoming insoluble. Potential solutions are:
  • Induce at a lower temperature (as low as 15°C overnight)
  • Reduce IPTG concentration to 10–40 µM
  • Induce earlier in growth phase (OD600 = 0.3 or 0.4) and harvest cells earlier.
  • Create an expression construct where the target protein is fused to Maltose Binding Protein (NEB #E8200).