TEV protease FRET activity assay kit (TEV protease activity kit)

Introduction

TEV protease activity assay kit for quantitative determination and monitoring of TEV protease specific activity based on fluorescent energy transfer (FRET). For quality control of TEV protease activity.

Description

The TEV protease fluorescent activity assay uses protein substrate based on fluorescent energy transfer (FRET). The principle of FRET relies on energy transfer of excitation energy of a donor fluorophore to a nearby acceptor fluorophore. The 54 kDa FRET substrate protein is composed by two fluorescent proteins green 26 kDa and red 28 kDa 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 range, the excitation range is 490-515 nm). The TEV protease fluorescent activity assay is suitable for precise protease activity measurements, monitoring or high-throughput screening of TEV protease variants, substrate specificity measurements, etc. All components are manufactured in certified laboratory environment and could be used in GMP certified downstream processes. Recombinant proteins (TEV protease and FRET TEV protease substrate) are purified by affinity chromatography, size exclusion chromatography and desalting.

Application

Activity assay for precise measurements of different lots and types of TEV protease. For 10 reactions and 2 calibration curves.

Purification method

Affinity chromatography, size exclusion chromatography, desalting.

Formulation

FRET TEV protease substrate (0,5 mg/ml; 1,5 ml); reference TEV protease (10 kU/ml, 50 μl); Protease reaction buffer (1 ml); DTT (1M DTT; volume 100 μl)

Specificity

TEV protease recognition sequence.

Storage

-20˚C