Protein Expression Using NiCo21(DE3) (C2529)

Protocol

1. Transform expression plasmid into NiCo21(DE3). Plate on antibiotic selection plates and incubate overnight at 37°C.
2. Resuspend a single colony in liquid culture with antibiotic to produce a starter culture. Inoculate starter culture at a 1:100 dilution into expression media containing antibiotic.
3. Incubate at 37°C with shaking until OD₆₀₀ reaches 0.4–0.8.
4. For most vector systems, induce expression with 40 or 400 µM IPTG for 3 hours at 37°C, 5 hours at 30°C or overnight at 16°C or 23°C.