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In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
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In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity

Article DOI: 10.3791/56629-v • 09:45 min • January 29th, 2018
January 29th, 2018 •
1, 2, 2,3,4,5, 1,6
1Institute of Microbiology and Immunology, National Yang-Ming University, 2UC Davis Cancer Center, University of California, Davis, 3Department of Biochemistry and Molecular Medicine, University of California, Davis, 4Institute for Translational Medicine, College of Medical Science and Technology, Taipei Medical University, 5Division of Molecular and Genomic Medicine, National Health Research Institutes, 6Center for Infectious Disease and Cancer Research, Kaohsiung Medical University

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Summary
Automatic Translation

Unlike ubiquitin ligases, few E3 SUMO ligases have been identified. This modified in vitro SUMOylation protocol is able to identify novel SUMO E3 ligases by an in vitro reconstitution assay.

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In Vitro SUMOylation Assay SUMO E3 Ligase Activity Protein Of Interest SUMOylated Protein SUMO E3 Ligase Identification SUMOylation Field SUMO E3 Ligase Specificity Tag K-bZIP Purification Antibody-tagged Magnetic Beads Cellular Lysate Suspension Mixer Centrifuge Wash Captured Proteins Lysis Buffer
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