Metadata |
datasetIdentifier | PASS01115 |
datasetType | SRM |
submitter | Victor Faundez <vfaunde@emory.edu> |
submitter_organization | Emory University |
lab_head_full_name | Victor Faundez |
lab_head_email | vfaunde@emory.edu |
lab_head_organization | Emory University |
lab_head_country | United States |
datasetTag | MSB-3677-3720-3747 |
datasetTitle | Genealogical Proteomics Menkes Disease |
publicReleaseDate | 2018-01-01 00:00:00 |
finalizedDate | |
summary | Triple SILAC labeling of human fibroblasts from families affected by Menkes disease |
contributors | Victor Faundez |
publication | Rare Disease Mechanisms Identified by Genealogical Proteomics of Copper Homeostasis Mutant Pedigrees. Zlatic et. al. Submitted |
growth | Seven passages in SILAC media |
treatment | |
extraction | Detergent Cell Extract |
separation | Gel digests were analyzed by nano LC/MS/MS with a Waters NanoAcquity HPLC system |
digestion | Gel pieces were processed using a robot (ProGest, DigiLab) with the following protocol:
· Washed with 25mM ammonium bicarbonate followed by acetonitrile.
· Reduced with 10mM dithiothreitol at 60°C followed by alkylation with 50mM iodoacetamide at RT.
· Digested with trypsin (Promega) at 37°C for 4h.
· Quenched with formic acid and the supernatant was analyzed directly without further processing. |
acquisition | data-dependent mode, with MS and MS/MS performed in the Orbitrap at
70,000 FWHM resolution and 17,500 FWHM resolution, respectively. The fifteen most abundant ions were
selected for MS/MS. |
informatics | MaxQuant software 1.4.1.2 |
instruments | ThermoFisher Q Exactive |
species | Human |
massModifications | Oxidation (M), Acetyl (Protein N-term), 13C6/15N2 (K), 13C6/15N4 (R), 4H2 (K), 13C6 (R) |