Figure 7From: Human Nek6 is a monomeric mostly globular kinase with an unfolded short N-terminal domainThermal shift denaturation of recombinant hNek6 variants fused to a 6xHis tag. The graph shows representative thermal shift data (markers) and fitted curves (lines). Proteins were buffered in 10 mM HEPES pH 7.5, 150 mM NaCl and assayed at a final concentration of 2.0 μM in 25 μL volume. SYPRO-Orange (Molecular Probes) was added as a fluorescence probe at a dilution of 1 in 1000. Temperature was raised with a step of 1°C per 1.0 min from 10°C to 85°C and fluorescence readings were taken at each interval. (wt: wild-type, D: dephosphorylated).Back to article page