Products/Services Used | Details | Operation |
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Proteins, Expression, Isolation and Analysis | The reaction products, along with matriptase alone or buffer alone controls, were first denatured by boiling at 95oC for 10 min, then loaded onto a 12% polyacrylamide gel (GenScript) and subjected to electrophoresis for protein fragment separation. | Get A Quote |
Dysregulated activity of the protease matriptase is a key contributor to aggressive tumor growth, cancer metastasis, and osteoarthritis. Methods for the detection and quantification of matriptase activity and inhibition would be useful tools. To address this need, we developed a matriptase-sensitive protein biosensor based on a dimerization-dependent red fluorescent protein (ddRFP) reporter system. In this platform, two adjoining protein domains, connected by a protease-labile linker, produce fluorescence when assembled and are nonfluorescent when the linker is cleaved by matriptase. A panel of ddRFP-based matriptase biosensor designs was created that contained different linker lengths between the p... More