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protein in phosphate buffer precipitate out with acetonitrile


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#1 mofeifei

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Posted 21 December 2009 - 05:42 PM

I want to quantify my protein by RP-HPLC. Usually my protein is just in water solution and I mix it with acetonitrile before loading onto the column. And it's fine. But recently my protein is in phosphate buffer. so when I mix my protein in phosphate buffer with acetonitrile, the protein would precipitate out. What should I do?

Thanks a lot!!

#2 mdfenko

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Posted 22 December 2009 - 07:30 AM

what is the pH of the phosphate buffer (sodium or potassium)? is it near the pI of the protein?

what is the concentration of acetonitrile? concentrated acetonitrile (ie high percentage or pure) can precipitate proteins.
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