Dear all,
I am trying to purify recombinant prion protein (PrP).
The chromatogram during gel permeation chromatography shows that nearly half of my protein exists in the fom of dimers.
Running buffer consists of 0,1 M citrate/phosphate, 0,2 M arginine and 50 ml KCl.
What can I do to minimize dimerization and increase monomeric yield? Are there any compounds I can add to the running buffer during gel filtration to prevent dimerization?
Thank you for any help!
Kind regards,
Peter
Hi Peter, If you see your protein in denaturing SDS- PAGE as a monomer, try to add 1-5 mM mercapto ethanol, moreover if yor protein has impaired cysteins. Good luck
Don't know if you alkylated the reduced protein gel sample with NEM or IAA as there are examples of in-situ oxidation during the gel run, hu IgG1 and GDF 15 are examples. Also NuPage reducing gel system can work.
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