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Hi All, I am trying to design a 1D 31P pH titration experiment for a protein. I have been trying to identify a good buffer system that covers a broad pH range. The protein is very stable so I can push the pH beyond the pH 3.6-9 range that you get with acetate/Tris-acetate. I've been searching through the literature, but I can't seem to find an alternative to the Tris-acetate buffer system (though I may be making my google-fu too narrow). Is anyone aware of a different buffer system for pH titrations of protein samples? Thanks for your time. |
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Do I understand you correctly that you're trying to find a single buffer system that spans a wide pH range? To do that, you'll definitely need to consider buffers with polyprotic acid components. However, why limit yourself to a single buffer composition? It's very common to screen the spectra of your protein in a wide variety of buffers to see which yield high quality spectra. As a starting point, take a look at this web page from the University of Buffalo: http://www.nmr2.buffalo.edu/nesg.wiki/Buffer_optimization There they give series of common buffers that span a large pH range. It's easy enough to buffer-exchange your protein and then record a quick spectrum. Since you're doing 31P NMR, simply exclude those buffers that contain phosphorous if you don't want to worry about extra signals appearing in your spectrum. good luck. |
