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Chapters
- 00:10Introduction
- 00:42Expression and Purification of Target Protein and Tagged Ubiquitin
- 03:15Immunoprecipitation
- 04:51SDS-PAGE and Immunostaining
- 05:48Detection of Protein Ubiquitination in vitro
- 07:02Representative Results
- 08:22Conclusion
Ubiquitination is a key posttranslational modification carried out by a set of three enzymes. Mutations of genes involved in this modification are associated with many different human diseases. Here, we describe protocols to detect protein ubiquitination in cultured cells in vivo and test tubes in vitro.
Tags
Protein UbiquitinationPosttranslational ModificationUbiquitin-activating Enzyme E1Ubiquitin-conjugating Enzyme E2Ubiquitin Ligase E3Deubiquitylating EnzymesProtein StabilityCellular LocalizationBiological ActivityGene MutationsHuman DiseasesCancerNeurodegenerationMetabolic DisordersPathogenesisDetection ProtocolsCultured Cells In VivoTest Tubes In VitroSumolyationNeddylation
