Lct premier tof mass spectrometer

Example 44

Mass spectrometry was used to analyze covalent modification of protein by selected inhibitors. Recombinant human PAD4 was used at 4 mg/ml in 20 mM Tris pH7.6, 400 mM NaCl, 5 mM TCEP. Where applicable, buffer was supplemented with 5 mM CaCl₂ to determine Calcium sensitivity of modification. Inhibitors were dissolved in DMSO at final concentrations of 0.2 mM to 5 mM and incubated with protein at 27° C. for 16 hours prior to analysis. Control experiments were performed with protein and DMSO in absence of inhibitor. Samples were centrifuged for 15 seconds at 10000 rpm at room temperature immediately prior to analysis using a Waters LCT-Premier TOF mass spectrometer, using a mobile phase from 5% to 80% acetonitrile in water supplemented by 0.1% formic acid.

Inactivation Kinetics

Example 18

Mass spectrometry was used to analyze covalent modification of protein by selected inhibitors. Recombinant human PAD4 was used at 4 mg/ml in 20 mM Tris pH7.6, 400 mM NaCl, 5 mM TCEP. Where applicable, buffer was supplemented with 5 mM CaCl₂) to determine Calcium sensitivity of modification. Inhibitors were dissolved in DMSO at final concentrations of 0.2 mM to 5 mM and incubated with protein at 27° C. for 16 hours prior to analysis. Control experiments were performed with protein and DMSO in absence of inhibitor. Samples were centrifuged for 15 seconds at 10000 rpm at room temperature immediately prior to analysis using a Waters LCT-Premier TOF mass spectrometer, using a mobile phase from 5% to 80% acetonitrile in water supplemented by 0.1% formic acid.