Journal
/
Biologie
/
In Vitro Analysis of E3 Ubiquitin Ligase Function
Journal JoVE
Biologie
Un abonnement à JoVE est nécessaire pour voir ce contenu.  Connectez-vous ou commencez votre essai gratuit.
Journal JoVE Biologie
In Vitro Analysis of E3 Ubiquitin Ligase Function

In Vitro Analysis of E3 Ubiquitin Ligase Function

DOI:
10.3791/62393-v

•

06:06 min

•

May 14, 2021

•

, , ,
1Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (CECAD),University of Cologne, 2Center for Molecular Medicine Cologne,University of Cologne

Chapitres

  • 00:05Introduction
  • 00:43In Vitro Auto-Ubiquitylation Assay
  • 01:31In vitro Substrate Ubiquitylation Assay
  • 02:13Lysine Discharge Assay
  • 03:37Results: Analysis of the Ubiquitylation Products
  • 05:28Conclusion

Summary

Traduction automatique

Traduction automatique

The present study provides detailed in vitro ubiquitylation assay protocols for the analysis of E3 ubiquitin ligase catalytic activity. Recombinant proteins were expressed using prokaryotic systems such as Escherichia coli culture.

Tags

Keywords: E3 Ubiquitin LigaseIn Vitro AnalysisE2 Enzyme SpecificitySubstrate SelectionUbiquitin TransferAuto-ubiquitylation AssaySubstrate Ubiquitylation AssayLysine Discharge AssayRecombinant ExpressionEukaryotic ProteinsSDS-PAGEThermal CyclerMaster MixEDTAApyrase

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

check_url/fr/62393?article_type=v

Analysis of the Development of a Morphological Phenotype as a Function of Protein Concentration in Budding Yeast
check_url/fr/62393?article_type=v

Analysis of Schwann-astrocyte Interactions Using In Vitro Assays
check_url/fr/62393?article_type=v

In Vitro Analysis of PDZ-dependent CFTR Macromolecular Signaling Complexes
check_url/fr/62393?article_type=v

Analysis of Gene Function and Visualization of Cilia-Generated Fluid Flow in Kupffer’s Vesicle
check_url/fr/62393?article_type=v

siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells
check_url/fr/62393?article_type=v

Analysis of Nephron Composition and Function in the Adult Zebrafish Kidney
check_url/fr/62393?article_type=v

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein
check_url/fr/62393?article_type=v

Comprehensive Workflow for the Genome-wide Identification and Expression Meta-analysis of the ATL E3 Ubiquitin Ligase Gene Family in Grapevine
check_url/fr/62393?article_type=v

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
check_url/fr/62393?article_type=v

Analysis of Beta-cell Function Using Single-cell Resolution Calcium Imaging in Zebrafish Islets

Read Article