Check for insolubility - this is important because high expression and high production of protein can result in the target protein becoming insoluble. Potential solutions for this are: » Induce at lower temperatures (as low as 12-15°C overnight) » Reduce IPTG concentration to between 0.01 mM and 0.1 mM » Induce for less time (as little as 15 minutes) » Induce earlier in growth (OD600 = 0.3 or 0.4)