Orbitrap elite ms detector

Tissue sample homogenization and sonication were carried out by means of the Wheaton® 903475 Overhead Stirrer apparatus (Wheaton, Millville, New Jersey, USA) and Branson Sonifier 450 (Branson Ultrasonics, Danbury, USA), respectively. Total protein concentration was determined in duplicate by Bradford assay (Bio-Rad Laboratories, Hercules, California, USA) by means of a UV-Vis spectrophotometer (8453 UV-Vis Supplies, Agilent Technologies, Waldbronn, Germany) using BSA as the protein of reference. HPLC-ESI-MS/MS analyses were performed on an UltiMate 3000 RSLCnano System coupled to an Orbitrap Elite MS detector with EASY-Spray nanoESI source (Thermo Fisher Scientific). EASY-Spray columns 15 cm × 50 μm ID, PepMap C18 (2 μm particles, 100 Å pore size), and 15 cm × 75 μm ID, PepMap C18 (5 μm particles, 300 Å pore size) (Thermo Fisher Scientific), were used for bottom-up and top-down analyses, respectively, in coupling to an Acclaim PepMap 100 cartridge (C18, 5 μm, 100 Å, 300 μm i.d. × 5 mm) (Thermo Fisher Scientific).

Tissue homogenization and sonication were carried out by means of a Wheaton^® 903475 Overhead Stirrer apparatus (Wheaton, Millville, NJ, USA) and a Branson Sonifier 450 (Branson Ultrasonics, Danbury, CT, USA), respectively. Total protein concentration was determined in duplicate by Bradford assay (Bio-Rad Laboratories, Hercules, CA, USA) and UV–Vis spectrophotometer (8453 UV–Vis Supplies, Agilent Technologies, Waldbronn, Germany) detector using BSA as the protein of reference. For sample centrifugation, a thermostated centrifuge SL16 R (Thermo Fisher Scientific, Langenselbold, Germany) or Mini Spin (Eppendorf AG, Hamburg, Germany) were used as specified for sample treatment. HPLC–ESI–MS/MS analyses were performed on an UltiMate 3000 RSLCnano System (Dionex, Sunnyvale, CA, USA) coupled with an Orbitrap Elite MS detector with ESI or EASY-Spray nanoESI sources (Thermo Fisher Scientific), as specified elsewhere.

Tissue homogenization and sonication were carried out by means of Wheaton^® 903475 Overhead Stirrer apparatus (Wheaton, Millville, NJ, USA) and Branson sonifer 450 (Branson Ultrasonics, Danbury, CT, USA), respectively. Total protein concentration was determined in duplicate by Bradford assay (Bio-Rad Laboratories, Hercules, CA, USA) and UV-Vis spectrophotometer (8453 UV-Vis Supplies, Agilent Technologies, Waldbronn, Germany) detector using BSA as the protein of reference. For sample centrifugation, thermostated centrifuge SL16 R (Thermo Fisher Scientific, Langenselbold, Germany) and Mini Spin (Eppendorf AG, Germany) were used as specified for sample treatment. HPLC-ESI-MS/MS analyses were performed on UltiMate 3000 RSLCnano System (Dionex, Sunnyvale, CA, USA) coupled to Orbitrap Elite MS detector with ESI or EASY-Spray nanoESI sources (Thermo Fisher Scientific), as specified elsewhere.