X500b q tof mass spectrometer

Protein samples were analysed by LC/MS, using a Sciex X500B Q-TOF mass spectrometer coupled to an Agilent 1290 Infinity II HPLC. Samples were injected onto a POROS R1 reverse-phase column (2.1 x 30 mm, 20 μm particle size, 4000 Å pore size) and desalted. The mobile phase flow rate was 300 μL/min and the gradient was as follows: 0-3 min: 0% B, 3-4 min: 0-15% B, 4-16 min: 15-55% B, 16-16.1 min: 55-80% B, 16.1-18 min: 80% B. The column was then re-equilibrated at initial conditions prior to the subsequent injection. Buffer A contained 0.1% formic acid in water and buffer B contained 0.1% formic acid in acetonitrile.
The mass spectrometer was controlled by Sciex OS v.1.6.1 using the following settings: Ion source gas 1 30 psi, ion source gas 2 30 psi, curtain gas 35, CAD gas 7, temperature 300 °C, spray voltage 5500 V, declustering potential 80 V, collision energy 10 V. Data was acquired from 400-2000 Da with a 0.5 s accumulation time and 4 time bins summed. The acquired mass spectra for the proteins of interest were deconvoluted using BioPharmaView v. 3.0.1 software (Sciex) in order to obtain the molecular weights. The peak threshold was set to ≥ 5%, reconstruction processing was set to 20 iterations with a signal-to-noise threshold of ≥ 20 and a resolution of 2500.

Protein samples were analysed by LC–MS, using a Sciex X500B Q-TOF mass spectrometer coupled to an Agilent 1290 Infinity II HPLC. Samples were injected onto a POROS R1 reverse-phase column (2.1 mm × 30 mm, 20 µm particle size, 4,000 Å pore size) and desalted. The mobile phase flow rate was 300 μl min⁻¹ and the gradient was as follows: 0–3 min, 0% B; 3–4 min, 0–15% B; 4–16 min, 15–55% B; 16–16.1 min, 55–80% B; 16.1–18 min, 80% B. The column was then re-equilibrated at the initial conditions before the subsequent injection. Buffer A contained 0.1% formic acid in water and buffer B contained 0.1% formic acid in acetonitrile.
The mass spectrometer was controlled by Sciex OS v.1.6.1 using the following settings: ion source gas 1, 30 psi; ion source gas 2, 30 psi; curtain gas, 35; CAD gas, 7; temperature, 300 °C; spray voltage, 5,500 V; declustering potential, 80 V; collision energy, 10 V. Data were acquired from 400–2,000 Da with a 0.5 s accumulation time and 4 time bins summed. The acquired mass spectra for the proteins of interest were deconvoluted using BioPharmaView v.3.0.1 (Sciex) to obtain the molecular mass values. The peak threshold was set to ≥5%, reconstruction processing was set to 20 iterations with a signal-to-noise threshold of ≥ 20 and a resolution of 2,500.