Hi all, is there any way of how to improve the protein yield during induction?
I am using the E. coli BL21 (DE3), LB medium and IPTG induction system. But I need to improve the protein yield during induction.
Which medium do you recommend, or which trick - e.g. ethanol in the medium, heat shock procedure or any supplement in medium for do this.
I do not need to have more cells, I need to have the higher concentration of desired protein in the cell. Thank you very much.
Pavel: There is not a single protocol that can provide the best yield. Oftentimes, a specific protein needs to be expressed in a particular cell line which has been optimized in many aspects. One needs to screen among different cell lines of the BL21 series, and perform protein analysis for the best expressor. IPTG and/or methanol use may have some impact but has far less impact on yield. Good luck.
Thank you, my protocol is well optimized now, because I am able to obtain huge quantitiy of protein. But I want to obtain even more of it, so I am interesting about the methods how to do it...
Hi Pavel,
You can try a richer expression medium. I've had good results with Turbo Broth from AthenaES. They also sell a media optimization kit with 6 different media to compare.
http://www.athenaes.com/Expression.php
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