Samples were dissolved in 12 µL solvent (98% water, 2% acetonitrile, and 0.1% formic acid), out of which 2.5 µL were injected into the nanoLC-MS/MS system consisting of a Dionex Ultimate 3000 RSLC nanoLC (Dionex, Sunnyvale, CA, USA) coupled to Bruker Maxis II Q-TOF apparatus (Bruker Daltonik GmbH, Bremen, Germany). Peptides were trapped on an Acclaim PepMap100 C18 (5 µm, 100 µm × 20 mm, Thermo Fisher Scientific, Waltham, MA) column and then separated on an Acquity M-Class BEH130 C18 analytical column (1.7 µm, 75 µm × 250 mm Waters, Milford, MA) using a gradient ranging from 4% to 50% eluent B in 120 min. Solvent A was water + 0.1% formic acid; Solvent B was acetonitrile + 0.1% formic acid. Spectra were collected using a fixed cycle time of 2.5 s and the following scan speeds: MS spectra at 3 Hz, while collision-induced dissociation (CID) was performed on multiply charged precursors at 16 Hz for abundant ones, and at 4 Hz for low abundance ones. Internal calibration was performed at the beginning of every measurement by sodium formate clusters and data were automatically recalibrated using Compass Data Analysis 4.3 (Bruker Daltonik GmbH, Bremen, Germany).
Acquity m class beh130 c18
Manufactured by Waters Corporation
The Acquity M-Class BEH130 C18 is a high-performance liquid chromatography (HPLC) column designed for separating and analyzing a wide range of compounds. It features a hybrid organic/inorganic bonded stationary phase with a particle size of 1.7 μm and a pore size of 130 Å, providing efficient separations and high resolution. The column is suitable for use in a variety of HPLC applications, including pharmaceuticals, environmental analysis, and food and beverage testing.
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2 protocols using acquity m class beh130 c18
1
Nano-LC-MS/MS Peptide Separation and Identification
2
Quantitative Nanoflow LC-QTOF Proteomics
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Samples were dissolved in 12 µL solvent (98% water, 2% acetonitrile and 0.1% formic acid) out of which 6 µL was subjected to nanoLC-MS/MS analysis using a Dionex Ultimate 3000 RSLC nanoLC (Dionex, Sunnyvale, CA, USA) coupled to a Bruker Maxis II Q-TOF (Bruker Daltonik GmbH, Bremen, Germany) via CaptiveSpray nanoBooster ionization source. Peptides were separated on an Acquity M-Class BEH130 C18 analytical column (1.7 µm, 75 µm x 250 mm Waters, Milford, MA) using gradient elution (4-50% eluent B in 120 minutes) following trapping on an Acclaim PepMap100 C18 (5 µm, 100 µm x 20 mm, Thermo Fisher Scientific, Waltham, MA) trap column. Solvent A consisted of water + 0.1% formic acid, while Solvent B was acetonitrile + 0.1% formic acid. Spectra were collected using a fix cycle time of 2.5 sec and the following scan speeds: MS spectra were acquired at 3 Hz, while CID was performed on multiply charged precursors at 16 Hz for abundant ions and at 4 Hz for low abundance ones. Internal calibration was performed by infusing sodium formate and data were automatically recalibrated using the Compass Data Analysis software 4.3 (Bruker Daltonik GmbH, Bremen, Germany).
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