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Title: human serum fractionation/depletion methods, 16_clinprot_c8      
description:
16_clinprot_c8
privacy:
not applicable
aggregation:
instance of dataset
ID:
PAe000338
refinement:
curated
availability:
available
types:
mass spectrometry
strain:
Male, Serum, Sigma-Aldrich, catalog #H4522, lot #043K0502
name:
Human
description:
Treatment: ; Growth: ; Digestion: Ammonium bicarbonate (50 mM) was added to achieve a final methanol concentration of 20% and the samples were digested with Trypsin Gold (Promega, Madison, WI) at a protein to enzyme ratio of 50:1 (w/w) at 37°C for 6 h. The samples were dried in a SpeedVac and resuspended in 50 mM ammonium bicarbonate prior to LC-MS analysis.; Extraction: Human serum samples were denatured and reduced with 60% methanol and 10 mM dithiothreitol (DTT) at 60°C for 1 h and alkylated with 50 mM iodoacetamide (IAM) at room temperature in the dark for 30 min.; Separation: Bruker ClinProt C8 magnetic beads ClinProt purification reagent sets were obtained from Bruker Daltonics (Billerica, MA). The magnetic beads were shaken 20 times to achieve a homogeneous suspension. MB-HIC binding solution (10µL) and 5µL of human serum were mixed in a thin wall PCR tube. Magnetic beads (5µL) were added to the mixture. After 1 min, the tube was placed in a magnetic bead separator (MBS) for 20 s to separate the beads from the supernatant. The beads were washed with 100µL washing solution three times. For elution, 5µL 50% ACN was added to the beads and mixed thoroughly. After 1 min, the tube was placed in a MBS and the beads were separated from the elution solution at the wall of the tubes.
name:
LTQ
accessURL: ftp://ftp.peptideatlas.org/pub/PeptideAtlas/Repository/PAe000338/
storedIn:
Peptide Atlas
qualifier:
gzip compressed
format:
XML
accessType:
download
authentication:
none
authorization:
none
abbreviation:
ISB
homePage: http://www.systemsbiology.org/
ID:
SCR:011305
name:
Institute for Systems Biology
homePage: http://www.peptideatlas.org/
ID:
SCR:006783
name:
PeptideAtlas