Epidermal lysates were obtained as described previously (Descargues et al., 2005 (link)). Briefly, epidermis was homogenized in 1 M acetic acid using Precellys 24 (Bertin Technologies). After overnight extraction at 4 °C, soluble proteins were lyophilized and resuspended in MiliQ water (Millipore, Bedford, MA). After acetone precipitation the BCA protein assay kit (Pierce, Rockford, IL) was used to quantify protein concentration. The soluble fraction, 2 μg, was monitored for protease activity using the EnzChek Protease Assay Kit (Life Technologies), according to the manufacturer’s instructions; 5 μg of the soluble fraction was loaded onto Novex Zymogram (casein) Gels (Life Technologies) for electrophoresis. Gels were stained with Coomassie brilliant blue.
Novex zymogram casein gels
Manufactured by Thermo Fisher Scientific
Novex Zymogram (casein) Gels are laboratory equipment used for the detection and analysis of proteolytic enzyme activity. They provide a platform for the visualization of enzyme-mediated casein degradation, allowing for the identification and characterization of proteases.
Automatically generated - may contain errors
2 protocols using novex zymogram casein gels
1
Epidermal Protein Extraction and Activity
2
Epidermal Proteolytic Activity Assay
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