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Capitoli
- 00:00Titolo
- 00:45Introduction
- 01:17Preparation of the Bait and SILAC Control Strain
- 02:44Grinding of Cells
- 04:05Pre-clearing Cell Lysate
- 05:51Eluting From IgG Beads by TEV Protease Cleavage
- 06:45Trichloroacetic Acid (TCA) Precipitation
- 08:11EtOH/ Acetate Precipitation
- 09:00Sample Analysis by Western Blotting and/ or Silver Staining
- 09:30Representative Results/ Outcome
- 10:15Conclusion
Hier beschreiben wir eine neue quantitative Proteomik Verfahren zur Identifizierung von Protein-Komplexen in Saccharomyces cerevisiae. In dieser Studie haben wir die SILAC-Methode zusammen mit Affinitätsreinigung durch Tandem-Massenspektrometrie gefolgt, um mit hoher Spezifität zu identifizieren die Bindungspartner einer ER-Protein, Scs2p verwendet.
Tags
Protein ComplexesQuantitative ProteomicsS. CerevisiaeLipidsCellular MembranesCompartmentalizationIntracellular Signaling MoleculesNonvesicular RoutesMembrane Contact Sites (MCSs)Endoplasmic Reticulum (ER)Plasma Membrane (PM)Lipid-synthesizing EnzymesLipid TrafficER-PM MCSsYeast ModelEukaryotesScs2pMolecular PartnersGel ElectrophoresisIn-gel DigestionMass Spectrometry
