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  • Reappraisal of quantitative gel zymography for matrix metalloproteinases.

Reappraisal of quantitative gel zymography for matrix metalloproteinases.

Journal of clinical laboratory analysis (2014-03-22)
Tommaso Prescimone, Danika Tognotti, Chiara Caselli, Manuela Cabiati, Andrea D'Amico, Silvia Del Ry, Daniela Giannessi
ABSTRACT

The determination of matrix metalloproteases (MMPs) is relevant in many pathophysiological conditions, especially if associated with extracellular matrix remodeling; however, the results obtained are closely linked to the method used and are not directly comparable. The aim of this study was to perform a reappraisal of quantitative gel zymography technique for MMPs in human plasma, to use for comparison with commercially available ELISA and in those experimental conditions where the MMP active form needs to be revealed. The critical methodological parameters of zymography were checked and a comparison with a routinely used ELISA was performed. Sensitivity and reproducibility levels of zymography are suitable for detection of MMP-9 in human plasma, providing results closely related to those obtained by ELISA. Analytical parameters of zymography were suitable for detection of MMPs in human plasma. Quantitative zymography for MMPs is an alternative method for comparing the results of ELISA widely employed for MMP determination, thus reducing the discrepancies between laboratories regarding gelatinase assay.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Phenylmethanesulfonyl fluoride, ≥98.5% (GC)
Sigma-Aldrich
Phenylmethanesulfonyl fluoride, ≥99.0% (T)
Sigma-Aldrich
Matrix Metalloproteinase-9 human, recombinant, >90% (SDS-PAGE), buffered aqueous solution