Glutathione beads for GST tagged protein. Umar
On Sat, 27 Jun 2020, 6:56 pm John Newitt, <newit...@gmail.com> wrote: > On Jun 27, 2020, at 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. > > > > > When you say “affinity column”, are you referring to a Ni2+-affinity > column? > -John > > > ######################################################################## 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/
