Hi It is hard to give specific advice without knowing details about your protein. There are multiple reasons that may be responsible for it's behavior (fundamentally the simplest reason is that proteins by and large are not designed by Nature to be purified and handled) so the more you share with us the more we may be able to help you.
Artem On Sat, Jun 27, 2020, 3:15 AM Umar Farook <umarfaroo...@gmail.com> wrote: > 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 > ######################################################################## 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/
