Rabbit anti p erbb4

Western blot was performed as previously reported (Dong et al., 2020 (link)). Lumbar enlargement segments of the spinal cord were dissected and the dorsal and ventral horns were carefully separated under a stereo microscope. Tissues were homogenized by sonication in RIPA buffer containing protease inhibitors. Debris was cleared by centrifugation at 12,000 g for 10 min at 4°C. Samples were resolved on SDS-PAGE and transferred to nitrocellulose membranes, which were incubated in PBS (phosphate buffered saline) containing 0.1% Tween-20 and 5% BSA for 1 hr at room temperature (RT) before overnight primary antibody incubation at 4°C. After wash, membranes were incubated with HRP-conjugated secondary antibody in PBS for 1 hr at RT. After incubating with enhanced chemiluminescence, immunoreactive bands were visualized by LI-COR Odyssey infrared imaging system. Intensity of immunoreactive bands were quantitated with ImageJ (NIH), with β-actin or GAPDH as loading control. The following primary antibodies were used: rabbit anti-ErbB4 (#0618, 1:2,000, generously provided by Dr. Cary Lai) (Zhu et al., 1995 (link)), rabbit anti-p-ErbB4 (#4757, 1:200, Cell Signaling Technology), mouse anti-β-actin (#3700, 1:5,000, Cell Signaling Technology), and mouse anti-GAPDH (#97166 1:5,000, Cell Signaling Technology).