We are working on the fibrillation of lysozyme. Given the acidic condition (pH 2) of the protein solution (glycine buffer) for fibrillation, what should be buffer and its pH for ThT stock solution?
Should the buffers of protein and ThT solutions be identical?
Hi,
ThT best response pH is about 8 (have a look at the enclosed publication), you should try to find a pH were ThT have a the best response and were your lysozymes are stable, maybe not more than 5 ?
I would say the buffers and ThT solution doesent need to have identical pH, but be carefull to not increas brutally the pH of your proteine solution to avoid local aggregation.
Good luck
The Thiflavin T stock solution has be around neutral pH and also the measurement. When you add a small amount of your amyloid testing solution that is at pH 2 it should not affect the measurement. You can do the thioflavin T test at lower pH.
I have never worked at such low pH with ThT. The best way would be to put ThT in water or in the same pH of your buffer but the efficiency of the absorption and emission will change with respect to physiological pH. I would suggest you to give a look at this paper (DOI 10.1007/s00249-015-1019-8 ) in which ThT was tested in a wide pH range.
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