Mwco lters

All CSF samples were grinded under liquid nitrogen conditions, mixed with protein lysate (7M urea, 2M thiourea, 4% SDS, 40mM Tris-HCl, pH8.5), 1mM phenyl methyl sulfonyl uoride (PMSF), 2mM ethylene diamine tetraacetic acid (EDTA) and 10mM dithiothreitol (DTT). The mixtures were ultra-sounded on ice for 10min and then centrifuged at 12000g, 4°C for 30min to collect liquid supernatant. Equal amounts of proteins were treated by reductive alkylation.
Then, equal amounts of proteins were ultra ltrated using molecular weight cut-off (MWCO) lters (Millipore, USA) of 10kDa according to the manufacturer's recommendation. The ow-through from the lters containing peptide fractions was recycled, desalted, concentrated using C18 solid phase extraction (SPE) (Strata™-X, 33μm, 2g/20mL, Phenomenex), and nally lyophilized.

All CSF samples were grinded under liquid nitrogen conditions, mixed with protein lysate (7M urea, 2M thiourea, 4% SDS, 40mM Tris-HCl, pH8.5), 1mM phenyl methyl sulfonyl uoride (PMSF), 2mM ethylene diamine tetraacetic acid (EDTA) and 10mM dithiothreitol (DTT). The mixtures were ultra-sounded on ice for 10min and then centrifuged at 12000g, 4°C for 30min to collect liquid supernatant. Equal amounts of proteins were treated by reductive alkylation. Then, equal amounts of proteins were ultra ltrated using molecular weight cut-off (MWCO) lters (Millipore, USA) of 10kDa according to the manufacturer's recommendation. The ow-through from the lters containing peptide fractions was recycled, desalted, concentrated using C18 solid phase extraction (SPE) (Strata™-X, 33μm, 2g/20mL, Phenomenex), and nally lyophilized.

All CSF samples were grinded under liquid nitrogen conditions, mixed with protein lysate (7M urea, 2M thiourea, 4% SDS, 40 mM Tris-HCl, pH8.5), 1 mM phenyl methyl sulfonyl uoride (PMSF), 2 mM ethylene diamine tetraacetic acid (EDTA) and 10 mM dithiothreitol (DTT). The mixtures were ultra-sounded on ice for 10 min and then centrifuged at 12000 g, 4 °C for 30 min to collect liquid supernatant. Equal amounts of proteins were treated by reductive alkylation. Then, equal amounts of proteins were ultra ltrated using molecular weight cut-off (MWCO) lters (Millipore, USA) of 10 kDa according to the manufacturer's recommendation. The ow-through from the lters containing peptide fractions was recycled, desalted, concentrated using C18 solid phase extraction (SPE) (Strata™-X, 33 µm, 2 g/20 mL, Phenomenex), and nally lyophilized.