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  • A novel simple fluorometric protease assay for monitoring hydrolysis of proteins in real time
    Bahun, Miha ; Poklar Ulrih, Nataša
    Measuring the activity of proteases is essential for investigating both the physiological functions and commercial applications of these enzymes. In contrast to the numerous protease assays that are ... based on chromogenic or fluorogenic peptide substrates, there is a lack of approaches to monitor degradation of proteins in real time. Here we report a protease assay where SYPRO Orange is employed as a fluorogenic probe to follow proteolysis. The functionality of the assay was demonstrated with the two subtilases of varying thermostability, using four different protein substrates. The assay is compatible with a real-time PCR instrument which allows continuous fluorescence measurements in low-volume samples even at high temperatures. This makes the assay especially suitable for high-throughput characterization of thermostable proteases.
    Source: Analytical biochemistry. - ISSN 0003-2697 (Vol. 696, [article no.] 115688, Jan. 2025, str. 1-9)
    Type of material - article, component part
    Publish date - 2025
    Language - english
    COBISS.SI-ID - 212076803

    Link(s):

    https://www.sciencedirect.com/science/article/pii/S000326972400232X?via%3Dihub
    Repository of the University of Ljubljana – RUL
    DOI

source: Analytical biochemistry. - ISSN 0003-2697 (Vol. 696, [article no.] 115688, Jan. 2025, str. 1-9)
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