Pooled, mixed-gender human blood sample (Golden West Biosolutions, LLC., Lot# PS1000.104) was spotted on Whatman cards. The discs with dried blood spots were transferred to 96-well plates (1.1 mL well volume, Thermo). The proteins were solubilized in 9 M Urea, 30 mM tris(2-carboxyethyl)phosphine (TCEP; Thermo Fisher Scientific), and 200 mM Trizma (Sigma-Aldrich) by shaking at 1200 rpm for 60 min at 37 °C. The remainder of the protocol was automated on a Biomek i7 workstation (Beckman Coulter). The samples were alkylated in 50 mM iodoacetamide (IAA) and then diluted with 200 mM Trizma to a final volume of 567 μL, and trypsin (Promega) was added to a 1:25 protease to substrate ratio. The plate was incubated at 42 °C for 4 h with slight shaking at 120 rpm and digestion was quenched by acidification to 1% formic acid. Digested samples were automatically desalted on an Oasis 30 μm HLB 96-well plate (Waters) following the manufacturer’s protocol adapted for the Biomek i7 with the positive pressure module.45
Biomek i7 workstation
Manufactured by Beckman Coulter
The Biomek i7 workstation is a high-performance automated liquid handling system designed for a wide range of laboratory applications. It features a modular design, advanced robotics, and precision liquid handling capabilities. The Biomek i7 enables efficient and reliable sample processing, assay development, and other laboratory workflows.
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Lab products found in correlation
2 protocols using biomek i7 workstation
1
Automated Whole Blood Protein Digestion
2
Automated Sample Preparation for LC-MS Analysis
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All sample preparation steps (methods 1-3) were performed using the Biomek i7 Workstation (Beckman Coulter Life Sciences), including the previously described positive pressure extractor, the Microplate Centrifuge VSpin, and integrated special solutions. Samples were analyzed by injecting 10 µL of the sample into an LC/time-of-flight (TOF)-MS system (Agilent Technologies) with a flow rate of 0.5 mL/ min. To increase the lifetime of the reverse-phase column Zorbax Eclipse PAH (Rapid Resolution HT, 4.6 × 50 mm, 1.8 µm, 600 bar), a 1260 Infinity LC in-line filter (4.6 mm) was installed before the chromatographic system. In addition to this measure, regular flushing with methanol and acetonitrile was performed. The mobile phase consisted of 92% methanol with 0.1% formic acid and 8% water with 0.1% formic acid. To avoid peak carryover, the run time of the method was 20 min. A dual ESI in positive ionization mode was used. The system was calibrated according to the internal standard method in the range of 0.
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