I want to remove Urea from my protein (6kDa) sample. I did use :
1. Step down the Urea gradient (dialysis) in order to remove urea and refold my protein, but my protein still contains urea in the range of 200 micromoles.
2. Although I used SEC after step down dilution, urea was still present in my sample.
3. I tried ethanol precipitation, but the structure of my protein was disrupted as a result.
The misfolded protein structure influences my experimental results.
Is there a solvent that dissolves urea without disturbing the protein structure at the same time?
I would appreciate your help with finding the best way to remove urea while maintaining protein concentration.
Thank you in advanced.
Before dialysis, you could try centrifugation the protein sample at 11000rpm for 15min.
I really appreciate your response Adam B Shapiro Fatemeh Yaghoobiadeh
Desalt ur protein in desalting columns such of the Biorad columns or any brand in hand, desalting in columns are much easier & quicker than dialysis , then concentrate ur protein.
Hope this helps
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