Pro3C HRV-3C Protease FRET activity assay kit (HRV-3C protease activity kit)

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Introduction

HRV-3C protease activity assay kit for quantitative determination and monitoring of HRV-3C protease specific activity based on fluorescent energy transfer (FRET). For quality control of HRV-3C protease activity.

Description

The HRV-3C 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 LEVLFQ|GP. 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 HRV-3C protease fluorescent activity assay is suitable for precise protease activity measurements, monitoring or high-throughput screening of HRV-3C 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 (HRV-3C protease and FRET HRV-3C protease substrate) are purified by affinity chromatography, size exclusion chromatography and desalting.

Application

Activity assay for precise measurements of different lots and types of HRV-3C protease. For 10 reactions and 2 calibration curves.

Purification method

Affinity chromatography, size exclusion chromatography, desalting.

Formulation

FRET HRV-3C protease substrate (0,5 mg/ml; 1,5 ml); reference HRV-3C protease (10 kU/ml, 50 μl); Protease reaction buffer (1 ml); DTT (1M DTT; volume 100 μl)

Specificity

HRV-3C protease recognition sequence.

Storage

-20˚C