In Vitro Competitive 5' Adenosine Monophosphate-activated Protein Kinase (AMPK) Assay
6:13
In Vitro AMPK Activity Assay with a Site-specific Mutant
7:39
Results: In Vitro AMPK Assays with Competitive Oligopeptides or a Site-specific Mutant Protein
8:31
Conclusion
Transcript
The overall goal of this procedure is to simultaneously assess multiple potential phosphorylation sites and to identify sites for subsequent validation by phosphorylation site mutants. This method can help answer key questions in the biochemistry
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Peptide competition assays are widely used in a variety of molecular and immunological experiments. This paper describes a detailed method for an in vitro oligopeptide-competing kinase assay and the associated validation procedures, which may be useful to find specific phosphorylation sites.
