Molecular Profiling of Neuronal Regeneration

The distal end of the nerve, muscles, dorsal root ganglion cells, and brain (hippocampus/cortex) were harvested 4 weeks after the various treatment and proteins were extracted. The cell lysate of dorsal root ganglion cells after electrical stimulation were collected to determine the expression of synaptophysin, TNF-α, and NGF. Proteins (50 μg) were resolved by SDS–polyacrylamide gel electrophoresis and were transferred onto a blotting membrane [20 (link)]. After blocking with non-fat milk, the membranes were incubated with antibodies against S-100 (Neomarkers, 1:500 dilution), NF (Cellsignal, 1:1000 dilution), synaptophysin (Abcam, 1:500 dilution), TNF-α (Abcam, 1:1000 dilution), and NGF-R (1:1000, Abbiotec) overnight at 4 °C. The intensity of the protein bands was determined by a computer image analysis system (IS1000, Alpha Innotech Corporation, CA, USA).