I want to optimize the protein expression by inducing at a low temperature. I am just wondering what is the best induction time at 16 degree. I am using BL21 and 0.5 M IPTG. Thank you guys.
In my hands, 48h-72h top get maximal accumulation of the expressed protein, but I think that this depends of the protein nature.
At 16 deg, you could incubate for anywhere from 18-72 h. It is also dependent on the inducer concentration and the nature of protein.
Every situation is different, so do a small scale expression test and take time points monitor by either Coomasie or silver stained SDS-PAGE and identify the best time point for yourself. You can also vary IPTG concentration and temperature, OD at induction so as to identify the best over-expression condition.
I agree about varying IPTG concentration- I usually decrease the IPTG concentration at lower temps, and when I do overnight or longer inductions. The goal is to express your protein slowly, giving the bacteria time to fold the protein correctly and keep it soluble.
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