Optimizing soluble protein extraction and two-dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates
Authors: Fulgentini L., Marangoni R., Colombetti G.
Autors Affiliation: Istituto di Biofisica del CNR, Via Giuseppe Moruzzi 1, I-56124 Pisa, Italy; Dipartimento di Scienze della Vita e dell\’Ambiente, Università del Piemonte Orientale Amedeo Avogadro, Alessandria, Italy; Dipartimento di Informatica, Università di Pisa, Pisa, Italy
Abstract: An efficient protein extraction methodology is quite important for sample preparation and subsequent 2-D PAGE and MS analysis. Cell lysis is the first step in protein extraction and putification. Many techniques are available for cell disruption, including physical and detergent-based methods. Here, we report on a very fast and efficient detergent-free Tris-based method to extract the soluble fraction proteins of extremophile ciliates, comparing it with a detergent-based protocol. This comparison has been carried out by means of 2-D PAGE and subsequent MALDI-compatible silver staining of protein samples obtained from the intensely pigmented hypersaline ciliate Fabrea salina and the Antarctic hypotrich ciliate Euplotes focardii. Our results indicate that this fast and easy extraction method allows to obtain more clear crude extracts and more spot-abundant polyacrylamide gels.
Volume: 29 (11) Pages from: 2411 to: 2412
KeyWords: Ciliates; 2-d PAGE; Extraction protocol; Extremophiles; Soluble proteinsDOI: 10.1002/elps.200700838Citations: 3data from “WEB OF SCIENCE” (of Thomson Reuters) are update at: 2019-10-13References taken from IsiWeb of Knowledge: (subscribers only)Connecting to view paper tab on IsiWeb: Click hereConnecting to view citations from IsiWeb: Click here