Sequential Detection of Thermophilic Lipase and Protease by Zymography.

Methods Mol Biol

Centre for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, Universidad de Carabobo, Valencia, Venezuela.

Published: March 2018

Lipase and protease present in cell-free fractions of thermophilic Bacillus sp. cultures were analyzed by polyacrylamide gel (PAG) electrophoresis. After run, the gel is electrotransferred to another PAG copolymerized with glycerol tributyrate, olive oil, and gelatin. This multi-substrate gel was incubated first for lipase detection, until bands appeared, and then stained with Coomassie for protease detection. Advantages of this sequential procedure are the detection of two different enzyme activities on a single PAG, beside time and resource saving.

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http://dx.doi.org/10.1007/978-1-4939-7111-4_25DOI Listing

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