Five microliter of the prepared sample was injected onto a Kinetex 2.6 m 100 100 2.1 mm (Phenomenex, Torrance, CA, USA) using SIL-30 AC auto sampler (Shimazdu, Columbia, MD, USA) connected with a high flow LC-30AD solvent delivery unit (Shimazdu) and CBM-20 A communication bus module (Shimazdu) online with QTRAP 5500 (Sciex) operating in positive and negative ion mode
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2.14 Differential scanning calorimetry (DSC) Being a tool that provides a thermodynamic profile of protein unfolding/denaturation process by measuring the heat absorbed or released by a protein sample as its temperature is changed, differential scanning calorimetry (DSC) is different from the biophysical technique discussed so far, all of which provide information on some structural features of a protein, such as its secondary structure, flexibility, hydrodynamic dimensions, or solvent/protease accessibility
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