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How it works:
Two interacting SNAP-tag fusion proteins are separately labeled in cell lysate: one with biotin, one with a fluorophore. The biotinylated protein is immobilized on a streptavidin-coated microtiter plate. Binding of the other protein is measured by fluorescence intensity.
Advantages:
- Specific covalent labeling in lysate, no prior purification needed.
- Labeling and immobilization are directed, leaving the protein free to interact.
- Wide choice of fluorophores.
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FKBP-FRB interaction assay with fluorescent and biotin labeled SNAP-tag fusion proteins in a streptavidin coated microtiter plate. Typical rapamycin dose response curves for this assay demonstrating labeling with different fluorophores.
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