FRET TEV protease substrate (TEV fluorescent substrate)

Introduction

Substrate protein for quantitative determination and monitoring of TEV protease specific activity based on fluorescent energy transfer (FRET).

Description

The 60 kDa FRET substrate protein is composed by two fluorescent proteins linked with TEV protease recognition sequence ENLYFQ|SG. The substrate is specifically cleaved to fluorescent monomers, which results in quantitative decrease of fluorescent intensity at 580-650 nm (emission). The excitation range of the substrate is 490-515 nm. Various TEV isolates (0.1 ug/ul) were incubated for indicated times with the FRET substrate (0.1 ug/ul) at 30°C (picture on right).

Application

Protease activity control and monitoring. High-throughput screening of TEV protease variants. This substrate is manufactured in certified laboratory environment and could be used in GMP certified downstream processes.

Purification method

Affinity chromatography, size exclusion chromatography, desalting.

Formulation

0.5 mg/ml, 10mM Tris pH7,5, 50mM KCl

Specificity

TEV protease recognition sequence.

Storage

-80°C, aliquot to avoid repeated freezing and thawing.