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Chapters
- 00:00Title
- 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
Burada Saccharomyces cerevisiae protein kompleksleri tanımlamak için yeni bir nicel proteomik tekniği açıklar. Bu çalışmada, yüksek özgüllük ile bir ER protein, Scs2p bağlayıcı ortakları tanımlamak için tandem kütle spektrometresi tarafından takip yakınlık arıtma ile birlikte SILAC yöntemi kullandık.
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
