Digested peptides were identified using nanoflow liquid chromatography coupled with amaZon speed ion trap mass spectrometry (Bruker-Michrom, Inc.), as previously described (17 (link)). The peptides were concentrated and desalted on a 75-umid×200 mmC18 Easy-nLC™ column (Thermo Fisher Scientific, Inc.). The mass spectrometer was operated in positive ion mode with a CaptiveSpray ion source and a spray voltage of 1,500 v, dry temperature of 150°C, without nebulizer gas and a mass range between 400-1,400 m/z. The parameter was optimized at 922 m/z with an ion charge count target of 400,000. To elute peptides, 0.1% formic acid in 3% acetonitrile (solution A) and 0.1% formic acid in 97% acetonitrile (solution B) were used with the following conditions: 10-70% B at 0-70 min, 90% B at 70-75 min and 10% B at 75-90 min. The raw data were processed using Bruker compass version 1.4 (Bruker-Michrom, Inc.). DataAnalysisTM version 4.0 (Bruker-Michrom, Inc.) created Mascot compatible files (mgf) to facilitate Mascot database searches. Each sample was assessed three times.
Compass version 1
Manufactured by Bruker
Sourced in Germany
The Bruker compass version 1.4 is a lab equipment product designed for analytical applications. It provides core functionality for users, though a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using compass version 1
1
Peptide Identification via Nanoflow LC-MS
2
Nanoflow LC-MS/MS Peptide Analysis
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The nanoflow liquid chromatography (Thermo Fisher Scientific, Waltham, MA, USA) coupled with amaZon speed ETD (Bruker Daltonics, Germany) with a CaptiveSpray ion source was used for peptide analysis. The tryptic-digested peptide samples were injected into an Acclaim PepMap RSLC C18 column (75 µm i.d. x 150 mm). The separation was performed at 300 nL/min using 70 min of 1–50% acetonitrile gradient containing 0.1% formic acid. The mass spectrometer was operated in the positive ion mode with a spray voltage of 1,500 V, dry temperature of 150˚C, without nebulizer gas and mass range between 400‑1,400 m/z. The parameter was optimized at 922 m/z with ion charge count target of 400,000. The raw LC‑MS/MS data were processed using Bruker Compass version 1.4 (Bruker Daltonics, Germany). Each sample was analyzed in triplicate, providing three technical replicates per sample.
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