Abstract
Gelatin zymography, first described by Heussen and Dowdle in the 1980s, is a widely used valuable tool in research and diagnostics. The technique identifies gelatinases by the degradation of their preferential substrate as well as by their molecular weight (kDa). We here describe detailed methodology for the detection of pro- and active- forms of both MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in cells using norepinephrine-stimulated H9c2 cardiomyoblasts as model. An easy to follow step-by-step protocol has been carefully written for reliable results. We also suggest an acceptable method for quantification of gelatin zymograms.
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Acknowledgment
This work is supported by the research grant awarded to Dr. Vibha Rani by the Science and Engineering Research Board (SERB), Department of Science and Technology (DST), Government of India (EMR/2016/005914).
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Chhabra, A., Rani, V. (2018). Gel-Based Gelatin Zymography to Examine Matrix Metalloproteinase Activity in Cell Culture. In: Cal, S., Obaya, A. (eds) Proteases and Cancer. Methods in Molecular Biology, vol 1731. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7595-2_9
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DOI: https://doi.org/10.1007/978-1-4939-7595-2_9
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