Re: [ccp4bb] CCP4BB Digest - 4 May 2009 to 5 May 2009 (#2009-124)

2009-05-06 Thread Geoffrey Feld
Refold protein 8M Urea: Sanjiv, The recommendations for rapid dilution and trying 6M GdmCl are great recommendations. As an alternative, I had a similar problem years ago with a membrane-associated receptor domain that was mostly found in inclusion bodies after expression (I assume this is your

Re: [ccp4bb] CCP4BB Digest - 4 May 2009 to 5 May 2009 (#2009-124)

2009-05-06 Thread Dima Klenchin
I incubated my pelets in 8M urea for a couple of hrs at 37C, spun and collected the supernatant, and then refolded the protein over a Ni column, as I had a his tag protein. To do this, you load and wash the protein normally, but before eluting, run b-cyclodextrin (i think 0.1%, you'll have to