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netBRET Signal
- The net BRET (Bioluminescence Resonance Energy Transfer) signal is a measure of the transfer of energy from a bioluminescent donor molecule to a fluorescent acceptor molecule in a protein-protein interaction assay. BRET is a sensitive and non-invasive technique that can be used to study protein-protein interactions in living cells.
- In a typical BRET assay, the donor molecule (usually a luciferase enzyme) is fused to one protein of interest, and the acceptor molecule (usually a fluorescent protein) is fused to another protein of interest. When the two proteins interact, energy is transferred from the donor to the acceptor, resulting in an increase in the fluorescence signal of the acceptor.
- The net BRET signal is calculated by subtracting the background signal (measured in the absence of the donor or acceptor) from the total signal (measured in the presence of both the donor and acceptor). This provides a quantitative measure of the protein-protein interaction, with a higher net BRET signal indicating a stronger interaction.
- The net BRET signal is used to determine the affinity, stoichiometry, and kinetics of protein-protein interactions, and is a valuable tool for drug discovery and development, as well as for basic research in cell biology and biochemistry.