Western Blotting Characterization of Oligodendrocyte Markers

Tissue samples were prepared, run on SDS-PAGE gels and transferred to PVDF membranes as described^{34 (link)}. Transferred membranes were incubated with primary antibodies overnight at 4°C. Primary antibodies used were as follows: Rabbit anti-Olig2 (1:2,000 Millipore AB9610), Mouse anti-MBP (1:2,000 Covance SMI-99P-100), Mouse anti-MOG (1:3,000 Millipore MAB5680), Rabbit anti-Neurofilament H (1:10,000 Millipore AB1989), Rabbit anti-GDE2 (1:1000), Rabbit anti-PDGF receptor α (1:2,000 Cell Signaling Technology 3174), Mouse anti-Actin (1:10,000 Millipore MAB1501). After 1 hour incubation at room temperature with appropriate HRP-conjugated secondary antibodies, membranes were developed by film or by using a digital imaging system (KwikQuant, Kindle Biosciences).