Protein Expression Using BL21(DE3) (C2527)
Transform expression plasmid into BL21(DE3).
Plate on antibiotic selection plates.
Incubate overnight at 37°C.
Resuspend a single colony in 10 ml liquid culture with antibiotic.
Incubate at 37°C until OD600 reaches 0.4–0.8.
Induce with 4 or 40 µl of a 100 mM stock of IPTG (final concentration of 40 or 400 µM)  Induce for 3 to 5 hours at 37°C.
Check for expression either by Coomassie stained protein gel, Western Blot or activity assay.
Check expression in both the total cell extract (soluble + insoluble) and the soluble fraction only.
