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The labelled peptide samples were combined and fractionated off-line into 20 fractions by strong cation exchange chromatography (ICAT strong cation exchange cartridge; Applied Biosystems) at a flow rate of 50 µl/min.
The generated tryptic peptides were fractionated using off-line HPLC and each fraction was further analyzed by nano-LC-MS/MS.
The resulting mixture was washed with brine (4×50 mL), dried (MgSO4) and concentrated in vacuo to give an off-white residue that was fractionated on silica with EtOAc.
Serum was fractionated on hydrophobic columns.
After detergent treatment, erythrocyte membranes were fractionated on sucrose gradients.
Further murine and human samples were fractionated using an off-gel fractionator fitted with 24 cm IPG strips according to the manufacturer's instructions (Agilent Technologies).
Samples containing high-intensity peaks to be identified were fractionated using an off-gel fractionator fitted with 24-cm IPG strips according to the manufacturer's instructions (Agilent Technologies, Santa Clara, CA, USA).
The polymers were fractionated by fractional precipitation.
CSF peptides were fractionated into 30 fractions.
The gradient was then fractionated into 12 fractions [ 13].
To better understand how these miRNAs were secreted into milk and the potential mechanism of uptake of milk miRNAs by the suckling young, milk was fractionated by ultracentrifugation and size cut-off column filtration.
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