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== Melting point == | == Melting point == | ||
Thermofluor is a fluorescence-based thermal stability assay. | |||
Ku T, Lu P, Chan C, Wang T, Lai S, Lyu P, Hsiao N. (2009) Predicting melting temperature directly from protein sequences. Comput Biol Chem. 33, 445-450 at [http://dx.doi.org/10.1016/j.compbiolchem.2009.10.002]. | Ku T, Lu P, Chan C, Wang T, Lai S, Lyu P, Hsiao N. (2009) Predicting melting temperature directly from protein sequences. Comput Biol Chem. 33, 445-450 at [http://dx.doi.org/10.1016/j.compbiolchem.2009.10.002]. | ||
They have a list of 35 different proteins with their Tms with the references from where they obtained their data, and their Tm Index program is available at [http://tm.life.nthu.edu.tw/]. | They have a list of 35 different proteins with their Tms with the references from where they obtained their data, and their Tm Index program is available at [http://tm.life.nthu.edu.tw/]. | ||
A specific case which shows that Tm does not need to be high: the protein with PDB-ID 1ofc had a melting temperature of 37°C (from CD), which was supported by the fact that it did not express in E.coli at that temperature. | |||
At 20°C it expressed to about 60mg / (liter LB), could be concentrated to more than 100mg/ml, crystallised at room temperature and diffracted to 1.9A. The initial purification steps were done at 4°C. |