Dear colleagues, I wonder if there were a bit less controversial possibility. No matter if that was less efficient. Would there be an option of using human cell lines that do not origin from killed children? As far as I know, the HEK cells do. Sometimes, the science can be pretty cruel. I am sorry for opening of this topic on a crystallographic forum, but so far, nobody has convinced me that I am allowed to work with these cell lines from a humanity (and Christian) point of view. Best regards, Petr
From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> On Behalf Of David Briggs Sent: Saturday, January 25, 2020 7:49 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] protein expression in human cells 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 <CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK>> on behalf of Rezaul Karim <00001e364c8f16de-dmarc-requ...@jiscmail.ac.uk<mailto:00001e364c8f16de-dmarc-requ...@jiscmail.ac.uk>> Sent: Saturday, January 25, 2020 1:31:02 AM To: CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK> <CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK>> 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: reza...@yahoo.com<mailto:reza...@yahoo.com>, rez...@health.usf.edu<mailto:rez...@health.usf.edu> Phone: +1-954-937-8487 On Friday, January 24, 2020, 4:51:11 PM EST, Gloria Borgstahl <gborgst...@gmail.com<mailto:gborgst...@gmail.com>> 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 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
