Overview

  • Product name

    TEV Protease Activity Assay Kit (Fluorometric)
  • Detection method

    Fluorescent
  • Sample type

    Cell Lysate, Purified protein
  • Assay type

    Enzyme activity (quantitative)
  • Sensitivity

    50 ng/well
  • Assay time

    0h 30m
  • Product overview

    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

  • Notes

    TEV Protease (Tobacco Etch Virus protease, EC: 3.4.22.44) 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.

  • Platform

    Microplate reader

Properties

Images

  • 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.

Protocols

References

ab211109 has not yet been referenced specifically in any publications.

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