Results: Specific Polyubiquitylation of UXT-V2-HA Mediated by Fbxo7
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Conclusion
Transcription
This protocol allows to evaluate the ubiquitylation of a specific substrate by an E3 ligase. We know that the characterization of ligase-substrate interaction is crucial to understanding their function in cells. This technique does not require exp
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We provide a detailed protocol for a ubiquitylation assay of a specific substrate and an E3 ubiquitin-ligase in mammalian cells. HEK293T cell lines were used for protein overexpression, the polyubiquitylated substrate was purified from cell lysates by immunoprecipitation, and resolved in SDS-PAGE. Immunoblotting was used to visualize this post-translational modification.