Products/Services Used | Details | Operation |
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Mutagenesis Services> | ... NJ; QuikChange ® Lightning Site-Directed Mutagenesis kit, Stratagene, La Jolla, CA; and the mKate S158A variant gene (GenBank Accession # EU383029) flanked by BamHI and XhoI restriction endonuclease sites in the pUC57 vector was synthesized by GenScript USA, Inc ... | Get A Quote |
We describe here the preparation of ratiometric luminescent probes that contain two well-separated emission peaks produced by a sequential bioluminescence resonance energy transfer (BRET)-fluorescence resonance energy transfer (FRET) process. The probes are single soluble fusion proteins consisting of a thermostable firefly luciferase variant that catalyzes yellow-green (560 nm maximum) bioluminescence and a red fluorescent protein covalently labeled with a near-Infrared fluorescent dye. The two proteins are connected by a decapeptide containing a protease recognition site specific for factor Xa, thrombin, or caspase 3. The rates of protease cleavage of the fusion protein substrates were monitored by recording ... More