Hi Gloria, Another vote here for HEK 293 Expi or Freestyle.
The off-the-shelf transfection reagents are super expensive, but I make my own (happy to share protocols with anyone interested) and we normally get a few mgs of protein per litre of suspension culture -- certainly enough for crystallography. If budget is an issue, semi-stable transfection of adherent HEK293s is much cheaper but the turnaround from vector to protein is much slower (a few weeks rather than a few days), and growing up panels of mutants can be quite tedious. Cheers & good luck Dave -- Dr David C. Briggs Senior Laboratory Research Scientist Signalling and Structural Biology Lab The Francis Crick Institute London, UK == about.me/david_briggs ________________________________ From: CCP4 bulletin board <[email protected]> on behalf of Rezaul Karim <[email protected]> Sent: Saturday, January 25, 2020 1:31:02 AM To: [email protected] <[email protected]> Subject: Re: [ccp4bb] protein expression in human cells In our lab, we see Expi293F works very good. Thanks, Reza Md. Rezaul Karim Graduate student, PhD Program in Integrated Biomedical Sciences Morsani College of Medicine University of South Florida E-mail: [email protected], [email protected] Phone: +1-954-937-8487 On Friday, January 24, 2020, 4:51:11 PM EST, Gloria Borgstahl <[email protected]> wrote: Hello CCP4-ers, I was wondering what people have found to be the best human cell line expression system for making a large quantity of purified recombinant protein. Any information and protocols would be greatly appreciated. Happy 2020, Gloria ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1<https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2Fwebadmin%3FSUBED1%3DCCP4BB%26A%3D1&data=02%7C01%7C%7Cdd4cd6f62475425a3afa08d7a1364e92%7C4eed7807ebad415aa7a99170947f4eae%7C0%7C0%7C637155126940120399&sdata=XEh8QKGrpCzQSR7EFXkx9uwNaJQGeUEZz6dmh%2FUQgso%3D&reserved=0> ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1<https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2Fwebadmin%3FSUBED1%3DCCP4BB%26A%3D1&data=02%7C01%7C%7Cdd4cd6f62475425a3afa08d7a1364e92%7C4eed7807ebad415aa7a99170947f4eae%7C0%7C0%7C637155126940120399&sdata=XEh8QKGrpCzQSR7EFXkx9uwNaJQGeUEZz6dmh%2FUQgso%3D&reserved=0> The Francis Crick Institute Limited is a registered charity in England and Wales no. 1140062 and a company registered in England and Wales no. 06885462, with its registered office at 1 Midland Road London NW1 1AT ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
