Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 30 min
- Sample type: Cell Lysate, Purified protein
- Sensitivity: 50 ng/well
Product nameTEV Protease Activity Assay Kit (Fluorometric)
Sample typeCell Lysate, Purified protein
Assay typeEnzyme activity (quantitative)
Assay time0h 30m
TEV Protease Activity Assay Kit (Flurometric) (ab211109) provides a convenient method for detecting TEV (Tobacco Etch Virus) Protease activity in cell lysates from infected individuals, as well as from purified proteins. The assay is based on the ability of TEV Protease to cleave a synthetic Fluorescein-based peptide substrate to release fluorescein which can be easily quantified using a fluorescence microplate reader at Ex/Em = 490/560 nm.
This assay kit is simple, rapid and can detect as low as 50 ng TEV Protease activity in samples.
TEV protease assay protocol summary:
- add samples and standards to wells
- add reaction mix
- analyze with microplate reader in kinetic mode for 15-30 min
TEV Protease (Tobacco Etch Virus protease, EC: 188.8.131.52) is a cysteine protease that recognizes the cleavage site of Glu-Xaa-Xaa-Y-Xaa-Gln-(Gly/Ser) and cleaves between Gln and Gly/Ser. The optimal sequence is Glu-Asn -Leu-Tyr-Phe-Gln-Ser/Glycine (ENLYFQS/G). TEV Protease has high specificity and great stability and is active over a wide range of temperatures (4-37°C) with an optimal activity at 34°C.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 100 tests TEV Protease Assay Buffer 1 x 25ml TEV Protease Dilution Buffer 1 x 10ml DTT (1M) 1 x 100µl TEV Protease (Positive Control) 1 x 10µl TEV Protease Substrate 1 x 100µl 5-FAM Standard (100 μM in DMSO) 1 x 100µl
Typical 5-FAM standard curve (20-100 pmol).
Adjusted 5-FAM Standard curve (2-10 pmol), to be used when sample shows TEV protease activity.
Kinetics progress curves observed for increasing amounts of TEV Protease (positive control included in kit). Assays were performed following the kit protocol.
Calculated ΔRFU (after 10 min) for different amounts of TEV Protease from kinetic curves shown in Kinetics progress curves observed for increasing amounts of TEV Protease graph. Assays were performed following the kit protocol.
ab211109 has not yet been referenced specifically in any publications.