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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
