P97 micropipette puller

Data Collection data was collected on two different cIM QToFs, a prototype instrument at Waters Corporation, Wilmslow and a commercial instrument at University College London. Samples were directly infused using nano ESI (nESI) gold-coated capillaries produced in-house using a Flaming-Brown P97 micropipette puller and coated using a Quorum Q150RS sputter coater. For data collection parameters for CytC, see table S1, and for IAPP see table S7.

All direct infusion mass spectra were collected on a Waters Micromass LCT Premier equipped with a Z-Spray nESI source. Borosilicate nESI needles were pulled to approximately 3–8 μm tip i.d. on a Sutter Instruments P-97 micropipette puller and gold coated using a Quorum Technologies SC7620 sputter coater as previously described.^{11 (link)} Each acquisition was performed with the emitter tip approximately 6 mm away from the sample cone. To each solution, unless otherwise noted, 0.2% ammonium hydroxide was added to promote analyte deprotonation. Sample flow rates were estimated to be 20–50 nl/min for direct infusion experiments.^{11 (link)} Each protein was examined at 10 μM. Peptide and small molecule concentration in their respective solutions were 5 μM. Spray voltage was applied at – (1.25–1.35) kV and remained constant for each experiment performed. The source region was heated to 65°C for all experiments.