This is a new approach to observing protein refolding by a thermal process, thus avoiding the use of denaturants such as urea and guanidine. The mT-temperature jump is an accessory for Bio-Logic stopped-flow instruments. The solution containing unfolded protein is kept at a higher temperature and the refolding is initiated by diluting with cold buffer.
The amplitude of the temperature jump and mixing ratio's are software controlled. The jump can be in both directions (cooling or heating) which offers new investigation possibilities. The technique is compatible with all optical detection methods including circular dichroism, FT-IR and fluorescence anisotropy
Click here to contact usClassical temperature jump instruments using electrical and laser pulses can give time resolution of a few microseconds. However they are limited to 5°C amplitude, are unable to perform cooling jumps and the heat of dissipation limits kinetics to milliseconds. The mT-temperature jump provides millisecond resolution, an amplitude of 40°C and unlimited kinetic capabilities.
The cold jump from 85°C to 60° shown below demonstrates the performance of the Stopped-Flow thermal mixing.
Refolding of cytochrome-c, after a cold jump from 85°C to 60°C observed by fluorescence
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