Exact(3)
The flow rate was 1.0 ml/min on a Dionex-Ultimate 3000 system using a PDA-100 Photodiode Array Detector (Dionex GmbH, Idstein, Germany) with a Acclaim 120 C18 (250 mm × 4.6 mm, 5 μm, 120 Å) column.
After concentration on a Acclaim PepMap 100 trap column at a flow rate of 5 μl/min (75 μm x 2 cm, C18, 3 μm, 100 Å, Thermo Scientific) separation was performed by UHPLC (UltiMate 3000, Dionex) using a Acclaim PepMap RSLC column at a flow rate of 300 nl/min (75 μm x 150 mm, C18, 2 μm, 100 Å, Thermo Scientific).
Determination of the peptides was carried out using a acclaim pep map 100 C18 nano-column (75 μm × 15 cm, 3 μm) operated at 300 nL min−1 combined with ICP-MS detection and post column addition of Lu yielding a recovery of nearly 100%and4.9%9% precision.
Similar(56)
10 μl of sample (a total of 2 μg) in 1% (vol/vol) formic acid was loaded with a constant flow of 4 μl/min onto an Acclaim PepMap100 columnlumn (0.3 mm id × 5 mm, Dionex Corporation, Sunnyvale, CA).
On average 0.5 µg was loaded with a constant flow of 5 µl/min onto an Acclaim PepMap100 nanoViper C18 trap column (100 µm inner-diameter, 2 cm; Thermo Scientific).
On average 0.5 μg was loaded with a constant flow of 5 μl min 1 onto an Acclaim PepMap100 nanoViper C18 trap column (100 μm inner-diameter, 2 cm; Thermo Scientific).
The immunoprecpitate solution was then separated by a biocompatible Ultimate 3000 nanoLC system (Dionex, Thermo Scientific) equipped with an Acclaim PepMap300 C18 trap column (300 μm id x 5 mm cartridge, Dionex) and an Acclaim PepMap300 C18 analytical column (75 μm id × 150 mm long, Dionex) with column oven temperature set to 45°C.
The instrument was online coupled to an Ultimate 3000 RSLCnano System (Dionex, Idstein, Germany) equipped with an Acclaim PepMap RSLC, 75 μm × 25 cm, C18, 2 μm, and 100 Å column.
The acidified peptides were separated on an Acclaim PepMap 100 C18 trap and RSLC C18 column (Thermo Fisher Scientific, UK), using a nanoLC Ultra 2D plus loading pump and nanoLC as-2 autosampler (Eskigent, UK).
Samples were injected into an Acclaim Pepmap 100 c18 1.0 mm × 150.0 mm column (Thermo) using an Ultimate 3000 nano HPLC pump (Dionex) to generate a post-split flow rate of 3 μl/min.
The detergents were separated using HPLC and an Acclaim Surfactant Plus 3.0 μm column (Thermo Scientific , Inc.
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