Dear All, Sorry for an offtopic question, your suggestions are highly appreciated.
We have been working on iron sulfur cluster binding protein, which is usually expressed as a nice soluble protein expressed in BL21 cells but aggregated in the affinity column itself and unable to recover from it. We had made n number of truncations and fused to soluble tags such as MBP, but always ended up in large aggregates. Anyone has experience in working with iron-sulfur cluster binding protein before, please let us know the critical steps in purification of such proteins, whether you have completely done the expression, purification and crystallization in anaerobic conditions? or else changing the expression system to eukaryotic system such as Baculo or HEK 293T would help? Please share your valuable experience, thank you. -- Best Regards, Umar Farook ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
