ridhima Posted August 12, 2013 Share Posted August 12, 2013 I want to know in pET28a/BL21DE3 and also in pCold TF/BL21DE3 for how much time (hrs) after induction with IPTG culture should be kept and at which temperature so that soluble aggregates should not form? I kept at 18 degree for 18 hrs after induction in pET28a and at 15 degree for 24 hrs but there is formation of soluble aggregates in both as detected by Native PAGE . I read that if kept for long time protein forms aggregates or come in inclusion bodies. Link to comment Share on other sites More sharing options...
BabcockHall Posted August 20, 2013 Share Posted August 20, 2013 In my limited experience, each situation is different. Therefore, you will probably need to experimentally determine conditions. If your protein has a prosthetic group, sometimes the growth medium needs to be enriched a chemically related species. Some proteins, however, refold nicely from inclusion bodies. Link to comment Share on other sites More sharing options...
ridhima Posted August 20, 2013 Author Share Posted August 20, 2013 ok sir !!!! thanks!!!!!!!! Link to comment Share on other sites More sharing options...
Recommended Posts
Create an account or sign in to comment
You need to be a member in order to leave a comment
Create an account
Sign up for a new account in our community. It's easy!
Register a new accountSign in
Already have an account? Sign in here.
Sign In Now