Preparation of Cul1AMC•Rbx1, the FRET Donor Protein
4:42
Preparation of FlAsHCand1, the FRET Acceptor Protein
5:18
Testing and Confirmation of the FRET Assay
6:08
Measuring the Association Rate Constant
7:37
Measuring the Dissociation Rate Constant of Cul1•Cand1 in the Presence of Skp•F-box Protein
8:33
Results: Dynamics Of Protein Complexes
10:30
Conclusion
副本
Our method helps study the kinetics of a protein complex formation. It describes how tight a protein complex is, and if the protein complex formation is interfered by other proteins. This technique enables studying protein-protein interaction in q
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Protein-protein interactions are critical for biological systems, and studies of the binding kinetics provide insights into the dynamics and function of protein complexes. We describe a method that quantifies the kinetic parameters of a protein complex using fluorescence resonance energy transfer and the stopped-flow technique.