SensoLyte ® 520 TEV Activity Assay Kit *Fluorimetric*
Tobacco Etch Virus (TEV) proteinase is widely used to remove fusion tags from recombinant proteins. It recognizes specific sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly and cleaves between Glutamine and Glycine residues. TEV protease is active even at 4 °C, making it the protease of choice for fusion proteins or tags removal. The SensoLyte® TEV Protease Assay Kit uses TEV specific Glu-Asn-Leu-Tyr-Phe-Gln-Gly substrate labeled with 5-FAM dye and QXL® 520 quencher. Proteolytic cleavage of this quenched 5-FAM-QXL® 520 conjugate yields a brightly green fluorescence, which can be continuously monitored at Ex/Em= 490/520 nm. Increase in fluorescence intensity is directly proportional to the TEV protease quantity/activity.
The kit contains: • 5-FAM/QXL® 520 TEV substrate, Ex/Em=490/520 nm upon cleavage• Assay buffer • Recombinant TEV Protease• DTT • Fluorescence reference standard for calibration • A detailed protocolKit size: 100 assays (96-well plate) Figure 1. Proteolytic cleavage of 5-FAM-QXL® 520 labeled TEV substrate. Fluorescence signal was measured with a fluorescence microplate reader (Flexstation 384II, Molecular Devices) at Ex/Em=490 nm/520 nm.