TEV Protease Activity assay kit utilizes the ability of TEV Protease to cleave a synthetic Fluorescein-based peptide substrate to release fluorescein which can be easily quantified using a fluorometer or fluorescence microplate reader (Ex/Em = 490/560 nm). This assay kit is simple, rapid and can detect TEV Protease activity as low as 50 ng/well in viral lysates and purified protein samples.
TEV Protease (EC: 220.127.116.11, Tobacco Etch Virus protease) is a cysteine protease that recognizes the cleavage site of Glu-Xaa- Xaa-YXaa-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.
Detection of TEV Protease activity in viral lysates Determine activity of purified TEV Protease.
Store kit at -20°C, protected from light. Briefly centrifuge small vials at low speed prior to opening. Read entire protocol before performing the assay.TEV Protease Assay Buffer: Add 100 µl of 1 M DTT to the TEV Protease Assay Buffer. Mix well. Store at -20°C. Bring to room temperature before use.TEV Protease (Positive Control): Aliquot and store at -80°C. Avoid repeated freeze/thaw.
TEV Protease Assay Buffer: 25 mlTEV Protease Dilution Buffer: 10 ml1 M DTT: 100 μlTEV Protease (Positive Control): 10 μlTEV Protease Substrate: 100 μl5-FAM Standard (100 µM in DMSO): 100 μl
Fluorescence (Ex/Em = 490/560 nm)
Compatible Sample Types:
Viral Lysates, purified protein
Features & Benefits:
• Simple, rapid assay• Can detect as low as 50 ng/well of TEV Protease activity• Fluorometric Detection