| Assay Method Information | |
| | Z-LYTE Kinase Assy |
| Description: | The Z-LYTE assay (Invitrogen Corporation) employs a FRET-based, coupled-enzyme format and is based on the differential sensitivity of phosphorylated and non-phosphorylated peptides to proteolytic cleavage. A site-specific protease recognizes and cleaves nonphosphorylated peptides. Phosphorylated peptides exhibit suppressed cleavage. Cleavage disrupts FRET between the donor (coumarin) and acceptor (fluorescein) fluorophores on the peptide, whereas uncleaved, phosphorylated peptides maintain FRET. The ratio of donor emission to acceptor emission after excitation of the donor fluorophore at 400 nm, quantitates reaction progress. |
| Affinity data for this assay | |
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