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Biochemistry

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JoVE Journal Biochemistry
Evaluering av substrat Ubiquitylation av E3 Ubiquitin-ligase i pattedyrcellelysater
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Evaluering av substrat Ubiquitylation av E3 Ubiquitin-ligase i pattedyrcellelysater

Article DOI: 10.3791/63561-v • 09:47 min • May 10th, 2022
May 10th, 2022 •
*1, *2, 1, 1,2
1Department of Genetic and Evolution, Federal University of São Carlos, 2Department of Biochemistry and Immunology, Faculty of Medicine of Ribeirão Preto, University of São Paulo
* These authors contributed equally

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Vi tilbyr en detaljert protokoll for en ubiquityleringsanalyse av et bestemt substrat og en E3 ubiquitin-ligase i pattedyrceller. HEK293T-cellelinjer ble brukt til proteinoverekspresjon, det polyubiquitylerte substratet ble renset fra cellelysater ved immunprecipitasjon og løst i SDS-PAGE. Immunoblotting ble brukt til å visualisere denne posttranslasjonelle modifikasjonen.

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