HPLC analyses were performed using an Ultimate 3000 SD System (Thermo Fisher Scientific, Waltham, MA, USA) coupled to a GabiStar detector (Raytest, Straubenhard, Germany). Compound were separated with a size exclusion column, XBridge protein BEH 200 Å SEC 3.5 μm, dimension 7.8 × 300 mm (Waters, Baden-Dättwil, Switzerland). Elution was performed using phosphate buffer pH 6.8 (1 mL/min) as mobile phase and was monitored via absorbance at 220/280 nm or γ detection.
Ultimate 3000 sd system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Ultimate 3000 SD System is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It features a modular design, allowing users to configure the system to meet their specific requirements. The system includes a solvent delivery module, an autosampler, a column compartment, and a variety of detection options, such as UV-Vis and charged aerosol detection. The Ultimate 3000 SD System is capable of delivering accurate and reproducible results, with precise flow control and temperature regulation.
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Lab products found in correlation
3 protocols using ultimate 3000 sd system
1
Size Exclusion HPLC Analysis
2
High-Performance Liquid Chromatography Analysis
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As described in Delage et al. [19 (link)], HPLC analyses were done using an Ultimate 3000 SD System (Thermo Fisher Scientific, Waltham, MA, USA) and a GabiStar radiodetector (Elysia-Raytest GmBH, Straubenhard, Germany). A size exclusion chromatography was performed using phosphate buffer pH 6.8 as solvent and a 200 kDa size exclusion column (XBridge protein BEH, Waters, Baden-Dättwil, Switzerland). Each chromatography profile was analyzed at 280 nm.
3
Kinetic Analysis of Inhibitor-βME Adduct Formation
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The tested compounds (1 mM) were reacted with
indicated concentrations of βME (Tables S2, S3, and S5 ) at 37 °C in a 4:1 DMSO/PBS (pH 7.4) mixture.
The loss of inhibitor and formation of inhibitor-βME adduct
was monitored by HPLC (Ultimate 3000SD System, ThermoFisher) at 20
min to 1 h intervals depending on reaction rates. Inhibitor-βME
adduct was identified by MALDI-ToF MS (Voyager-DeTM Pro measured in m/z; see the MALDI-MS Spectra of β-Mercaptoethanol
Adducts section in theSupporting Information ). The experiments were performed in triplicate. For further details,
see theSupporting Information .
indicated concentrations of βME (
The loss of inhibitor and formation of inhibitor-βME adduct
was monitored by HPLC (Ultimate 3000SD System, ThermoFisher) at 20
min to 1 h intervals depending on reaction rates. Inhibitor-βME
adduct was identified by MALDI-ToF MS (Voyager-DeTM Pro measured in m/z; see the MALDI-MS Spectra of β-Mercaptoethanol
Adducts section in the
see the
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