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Urea buffer: To remove cations or not

Urea magnesium calcium cations mixed-bed IEC

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#1 Edmond Ahgumon Chua

Edmond Ahgumon Chua

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Posted 01 May 2012 - 10:12 PM

Hi all,

I'm trying to refold my protein (ScFv) from inclusion bodies. First step would be to resolubilize the inclusion bodies with a 8M Urea buffer. However, I have also read somewhere that the Urea buffer needs to have cations like Magnesium and Calcium removed so refolding is not affected. Is this true?
Would using a mixed-bed IEC help in purification downstream?

This is also the first time I am trying to refold proteins and would appreciate any tips/tricks that I could follow to ensure successful refolding. I'm planning to try step-wise dialysis to get a higher refolding efficiency.

Thank you.





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