Hi,
I am working on the protein denaturation by circular dichroism. I have tried both GdnHCl and urea and it did not work well. I want to try stronger denaturants, such as GdnSCN and Thiourea. Has any one use one of these to study protein denaturation by circular dichroism. I found some papers that they used GdnSCN for denaturation assay by Trp fluorescence, not CD. it looks like that GdnSCN will cause large noise in CD spectra. I can't find much information about Thiourea denaturation. Thanks!
Although I have not used the two new compounds you mention for CD, I have had problems with high amounts of urea and GdnHCl in CD, but did manage to get some data with lower molarities of urea. And, I have also found that urea and GdnHCl do not interfere with UV/Vis of fluorescence spectra. I suspect that the Gdn with thiourea will not work with CD, but the thiourea may work at lower concentrations. My only suggestion with the latter is to subtract a thiourea blank and hopefully you can get meaningful data.
To add, also be aware of the quality (purity level) of the denaturants. Some grades of these reagents give much less background.
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