Publication Type:

Journal Article

Source:

Electrophoresis, Volume 32, Number 24, p.3621-3629 (2011)

URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-83755220047&partnerID=40&md5=599787f09a214c15b84b40feadc7cec6

Keywords:

acetone, alcohol, Alkanesulfonates, ammonium acetate, Animals, article, Brain Chemistry, Chemical precipitation, controlled study, detergent, Detergents, Electrophoresis, Ethanol, gel, intermethod comparison, methanol, myelin, myelin protein, Myelin Proteins, myelin sheath, polyacrylamide gel electrophoresis, precipitation, protein analysis, protein purification, proteome, proteomics, Rats, silver staining, solubility, solubilization, thermodynamics, two dimensional gel electrophoresis, Two-Dimensional, Wistar

Abstract:

Performing 2-DE of lipid-rich multilamellar membranes like myelin is a cumbersome task. However, for understanding its molecular organization and changes during diseases, identification of proteins of myelin is essential. Although the 2-D-proteomic approach of myelin has been employed to understand the myelin proteome, representation of myelin proteins in its entirety is still a challenge. 2-DE profiling of myelin proteins is very important for the detection of immuno-reactivity to myelin proteins from various biological fluids following Western blotting in diseases like multiple sclerosis. Here we developed a novel approach by exploiting the thermodynamic principles behind detergent-mediated solubilization of myelin membranes without any conventional processing of myelin involving precipitation of myelin proteins. We show that the addition of myelin to ASB-14-4 resulted in significant increase in protein representation of myelin in 2-DE compared with the addition of ASB-14-4 to myelin. Moreover, the number and resolution of spots are significantly higher in myelin to ASB-14-4 strategy than other strategies of myelin sample processing such as ASB-14-4 to myelin or ethanol or acetone or methanol-ammonium acetate precipitation of myelin proteins. In addition, the step involves no precipitation that selective removal of any proteins as a result of precipitation is nil and a qualitative representation of myelin proteins in a 2-D gel is achieved. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Notes:

cited By (since 1996)2

Cite this Research Publication

S. Nair, Xavier, T., Kumar, M. K. Satheesh, Saha, S., and Menon, K. N., “Exploitation of detergent thermodynamics in the direct solubilization of myelin membrane proteins for two-dimensional gel electrophoresis for proteomic analysis”, Electrophoresis, vol. 32, pp. 3621-3629, 2011.