Zymography has long been used for the detection of substrate-specific enzyme activity. In situ zymography (ISZ), an adaptation from the conventional substrate zymography, is a widely employed technique useful for the detection, localization, and estimation of enzyme–substrate interactions in tissues. Here, we describe a protocol to detect ‘in position’ matrix metalloproteinase (MMP) activity in cells utilizing H9c2 cardiomyoblasts as a model. This technique is primarily adopted from the method used for histological sections and is termed as ‘Cell in situ Zymography’. It is a simple, sensitive, and quantifiable methodology to assess the functional activity of an enzyme ‘on site/in position’ in cell culture.
CITATION STYLE
Chhabra, A., & Rani, V. (2017). Cell in situ zymography: Imaging enzyme–substrate interactions. In Methods in Molecular Biology (Vol. 1626, pp. 133–143). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7111-4_12
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