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Preparation of Cell Extracts by Cryogrinding in an Automated Freezer Mill
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Biologie
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Journal JoVE Biologie
Preparation of Cell Extracts by Cryogrinding in an Automated Freezer Mill

Preparation of Cell Extracts by Cryogrinding in an Automated Freezer Mill

DOI:
10.3791/61164-v

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14:00 min

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January 29, 2021

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, , , ,
1Department of Biomedical Sciences,Florida State University College of Medicine

Summary

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We describe a reliable method for the preparation of whole cell extracts from yeast or other cells using a cryogenic freezer mill that minimizes degradation and denaturation of proteins. The cell extracts are suitable for purification of functional protein complexes, proteomic analyses, co-immunoprecipitation studies and detection of labile protein modifications.

Tags

Yeast Cell LysisCryogrindingAutomated Freezer MillProtein ExtractionNative Protein IsolationMacromolecule ExtractionPhysical Lysis MethodsTemperature-sensitive LysisEnzymatic Lysis LimitationsChemical Lysis LimitationsFrench PressMortar And PestleBead Beater MillSonicationHeat GenerationProtein DenaturationProtein Degradation

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