Abstract
Zymography, the detection of proteolytic activities on the basis of protein substrate degradation, has been a technique described in the literature for at least in the past 50 years. In this study, we used substrate immersing zymography to analyze proteolysis of proteases. Instead of being directly added into a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS–PAGE) gel, the substrates were added into the immersing solution after electrophoresis. With substrate immersing zymography, some characters of proteases, such as enzyme forms, potential proteolytic activity, molecular weights, presence of complexes, and potentially active enzyme fragments in complex biological samples, can be determined.
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Acknowledgments
The work was supported by National Natural Science Foundation of China (31070061, 31370104), Hunan Provincial Natural Science Foundation of China (13JJ9001), and National Sparking Plan Project (2013GA770009).
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He, H., Li, H., Liu, D. (2017). In Situ Demonstration and Characteristic Analysis of the Protease Using Substrate Immersing Zymography. In: Wilkesman, J., Kurz, L. (eds) Zymography. Methods in Molecular Biology, vol 1626. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7111-4_19
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DOI: https://doi.org/10.1007/978-1-4939-7111-4_19
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