Abstract
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) provides high resolution separation of proteins and offers a powerful method for their identification and characterization. Since many myelin-specific proteins are highly basic, they cannot readily be analyzed by standard isoelectric focusing (IEF)-2D-PAGE that affords separation primarily in the isoelectric points (pI) range of 4-8. An alternative method, nonequilibrium pH gradient electrophoresis (NEPHGE)-2D-PAGE, can provide excellent resolution of highly basic proteins. In the present study, we have optimized the NEPHGE-2D-PAGE protocol for the analysis of myelin proteins with basic pIs, and provide a NEPHGE-2D-PAGE map based on size, pI, and immunoreactivity (Western blot) of myelin basic protein (MBP), 2', 3'-cyclic-nucleotide 3'-phosphodiesterase (CNP), myelin proteolipid protein (PLP), and its smaller spliced variant DM20, myelin/oligodendrocyte glycoprotein (MOG) and oligodendrocyte-specific protein (OSP). We have also demonstrated, by analyzing metabolically radiolabeled oligodendrocytes in culture at specific stages of the developmental lineage, the developmentally up-regulated expressions of several undefined, oligodendrocyte, basic membrane proteins during oligodendrocyte differentiation. We suggest that this approach offers an important tool for identifying and characterizing the plethora of uncharacterized myelin proteins.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
