Zymography pp 287-293 | Cite as
CTAB Zymography for the Analysis of Aspartic Proteases from Marine Sponges
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Abstract
Electrophoresis under denaturing conditions in the presence of SDS is a standard method for the protein and enzyme scientist. Nevertheless, there are special situations where this method may originate nonoptimal results. SDS may cause protein aggregation or precipitation. Beyond this, depending on the type of protein, some just do not resolve well or migrate abnormally in SDS gels. SDS, an anionic detergent, may be however substituted by a cationic detergent, like CTAB (cetyltrimethylammonium bromide), in order to solubilize and electrophorize proteins. CTAB electrophoresis allows the separation of proteins based on molecular weight and can be carried out at neutral or acidic pH. Here, we describe the development of a CTAB zymography method to analyze aspartic proteases from marine sponges, which present an abnormal high R f value when run in SDS-PAGE. The special feature of using CTAB is that it binds proteins, making them positively charged and thus migrating in the opposite direction compared to SDS-PAGE.
Key words
Aspartic proteases CTAB Cationic zymography ElectrophoresisNotes
Acknowledgments
We acknowledge partial funding from the Consejo de Desarrollo Científico y Humanístico de la Universidad de Carabobo (CDCH-2014). We kindly thank the donation of CTAB from Prof. Dr. C. Cabrele, University of Salzburg, Austria. Sponge samples were kindly provided by Prof. J.G. Rodríguez, Biology Department, FACYT, University of Carabobo.
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