The Protease-Glo™ Assay is a novel method to detect and measure protease activities using a genetically engineered firefly (Photinus pyralis) luciferase and represents one example of the GloSensor™ platform technology. The assay uses a circularly permuted firefly luciferase, the GloSensor™-10F protein, with a protease recognition site as the protease substrate. This assay system allows rapid generation of protease substrates through molecular cloning and coupled transcription/translation cell-free expression, thus enabling the facile evaluation of protease function. Oligonucleotides encoding a protease recognition sequence are designed and cloned into the GloSensor™-10F gene located on a linearized vector. The GloSensor™ protein containing the protease site of interest is then synthesized in a cell-free protein expression system and subsequently used as a protease substrate. Cleavage of the protease recognition sequence leads to activation of the GloSensor™ protein and light emission. The level of luminescence correlates to protease activity. The Protease-Glo™ Assay has the advantage of a bioluminescent readout, which provides easy quantitation, high sensitivity and wide dynamic range.
Visit the Protease-Glo™ Assay Design Toolto see how to generate your protease recognition site of interest in the pGloSensor™-10F Linear Vector and express the protein using cell-free translation.
Features - Benefits
Flexible:Use with P´ requiring proteases.
Avoid Fluorescent Background Problems:Physical and chemical features of luminescence overcome problems due to fluorescence interference.
Greater Sensitivity:Ease and dynamic range of luminescence.
Open Platform System:Create your own recognition substrates.
Interrogate Sequences:Excellent tool to determine optimal protease recognition sequences or effects of amino acid substitutions.
Web Application:Makes proper oligo design fast and easy; simply enter your amino acid sequence of interest.
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