Optimizing soluble protein extraction and two-dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates

An efficient protein extraction methodology is quite important for sample preparation and subsequent 2D-PAGE and mass spectrometry analysis. Cell lysis is the first step in protein extraction and purification. Many techniques are available for cell disruption, including physical and detergent-based...

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Bibliographic Details
Published in:ELECTROPHORESIS
Main Authors: FULGENTINI LORENZO, COLOMBETTI GIULIANO, MARANGONI, ROBERTO
Other Authors: Fulgentini, Lorenzo, Marangoni, Roberto, Colombetti, Giuliano
Format: Article in Journal/Newspaper
Language:English
Published: 2008
Subjects:
Antarctic
The Antarctic
Antarc*
Online Access:http://hdl.handle.net/11568/123023
https://doi.org/10.1002/elps.200700838
Description
Summary:An efficient protein extraction methodology is quite important for sample preparation and subsequent 2D-PAGE and mass spectrometry analysis. Cell lysis is the first step in protein extraction and purification. 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 2D-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.

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