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

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 Pro3C 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 54kDa FRET substrate protein is composed by two fluorescent proteins, green 26kDa and red 28kDa proteins linked with HRV-3C protease recognition sequence LEVLFQ|GP. The substrate is specifically cleaved to fluorescent monomers, which results in quantitative decrease of fluorescent intensity at 580-650nm (emission range, the excitation range is 490-515nm). The Pro3C HRV-3C protease fluorescent activity assay is suitable for precise protease activity measurements, monitoring or high-throughput screening of 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 (3C-protease and FRET 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 (800 U/ml, 50 μl); Protease reaction buffer (1 ml); DTT (1M DTT; volume 100 μl)

Specificity

HRV-3C protease recognition sequence.

Storage

-20˚C