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. 2015 Jul 23;24(20):5697–5710. doi: 10.1093/hmg/ddv286

Figure 5.

Figure 5.

Thermal stability of HNMT (WT) and HNMT (Gly60Asp) using Thermofluor®. The experiment was performed by mixing 5 μl of 1 mg/mL of HNMT (WT) or HNMT (Gly60Asp) with 5 μl 200-fold diluted SYPRO Orange and a buffer containing 50 mm Tris–HCl and 100 mm NaCl, pH 8.0, in a 98-well RT-PCR plate. Reaction mixtures were heated at 0.5°C/min from 20 to 95°C. A plot between d(fluorescence)/dT versus temperature of HNMT (WT) (red line) and HNMT (Gly60Asp) (blue line) is shown. A melting temperature of 62.3 ± 1.7 and 58.3 ± 1.1°C for WT and the Gly60Asp variant, respectively, was obtained.