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Journal / Biology / Budding Yeast Protein Extraction and Purification …
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Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications
 

Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

Article DOI: 10.3791/50921-v • 09:22 min • October 30th, 2013
October 30th, 2013 •
1, 1
1Integrated Science Center, Biology Department, The College of William & Mary

Kapitel

Summary
Automatic Translation

The preparation of high quality yeast cell extracts is a necessary first step in the analysis of individual proteins or entire proteomes. Here we describe a fast, efficient, and reliable homogenization protocol for budding yeast cells that has been optimized to preserve protein functions, interactions, and post-translational modifications.

Tags

Budding Yeast Protein Extraction Purification Function Interactions Post-translational Modifications Bead Beating Homogenization DNA RNA Proteins Metabolites Bead Mill Homogenizer Buffers Protein Expression Liquid Growth Media Inducible Promoters Cell Cycle Stage Synchronization Proteasome Function Inhibition Pelleting Resuspension Buffer Protease Inhibitors Phosphatase Inhibitors Freezing In Liquid Nitrogen Homogenization Cycles Bead-beating Incubation On Ice Centrifugation Filtration Whole Cell Extract (WCE) TCA Precipitation SDS-PAGE Analysis Western Blotting
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