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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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protein induction
Posted by: kcoghlan (IP Hidden, New member, 0)
Date: June 16, 2004 05:19PM
Hi there,
I am using the Pet21(d) vector in the E.coli strain BL21 Des (pLys) I am having a significant problem in growing up large batches of protein. I usually grow up 50 mls of cells at a time to see if the cells will produce my desired protein after induction with IPTG. I use a western blot to check to make sure the protein is being produced. I then use some of the cells from the same culture and upscale to 1 liter. However when I try to get protein from this larger batch I still have the same amount as in the the smaller batch which is a very tiny amount. I can't seem to concentrate this little bit of protein since it gets "lost" in subsequent purification. Does anyone have any ideas as to how I can scale up my protein production for research purposes? Thank you.
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