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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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Cytochrome P450 expression
Posted by: shirleyc (IP Hidden, New member, 2)
Date: November 2, 2005 06:38AM
Hi everyone,
I have tried to express a recombinant cytochrome P450 protein in E. coli BL21(DE3) using Prset as Expression vector. I was failed to purify the protein expressed in the system under both native (solubilizing the E. coli membrane by different combination of detergents) and denaturing condition (I have tried Urea, guanidine hydrochloride). Can anyone give me some suggestions on the purification of the cytochrome? Or is there anything that is crucial and critical for purifying these kinds of proteins? I know that most lab using carbon monoxide to quantify expressed cytochrome P450. However, my lab has never handled these kinds of proteins before. Therefore, we don't have any equipment that can handle carbon monoxide. So are there any other methods that can quantify the expressed cytochrome P450? Or can anyone give me some suggestions on how to set up those equipments for CO handling? And are there any regulation or safety measurement I must consider when I set up those equipments? I am looking forward to any suggestion or help! Thank you very much! Shirley
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