Super ecl detection kit

The detailed information for primary antibodies used in this study is described in Supplementary Table S4. For immunoblotting analyses, cells or tissue samples were lysed in RIPA buffer with 1 × protease inhibitors and phosphatase inhibitors (Bimake, #B14002 and #B15003, respectively). Quantification of protein concentrations was performed using the bicinchoninic acid assay kit (Yeasen, # 20201ES90). Equal amounts of protein extracts were separated by SDS-PAGE and transferred onto PVDF membranes (Millipore, #IPVH00010). The membranes were incubated with indicated primary antibodies and detected with Super ECL detection kit (Yeasen, # 36208ES76). Quantitation of immunoblotting bands was performed using ImageJ software, and expression levels of target proteins were normalized to those of internal control Vinculin.

Striatum tissues were lysed in RIPA Buffer (Beyotime, Shanghai) for protein extraction. After centrifugation at 12,000 rpm, 4°C for 15 min, the supernatant was collected and protein concentrations were determined with a PierceTM BCA Protein Assay Kit (Thermo Fisher Scientific). Proteins were separated by SDS-PAGE and transferred to PVDF membranes (Merck Millipore, Guangzhou, China). After blocking with 5% fat-free milk in 1XTBST (0.1% Tween20 in Tris-buffered saline), the blotted membranes were first incubated with primary antibodies (Supplementary Table 2) at 4°C overnight and then with HRP-conjugated secondary antibodies (Supplementary Table 2) at RT for 1 h, with TBST washing in between. Protein immunoreactivity was detected with a Super ECL Detection Kit (Yeasen Biotech, Shanghai), and the signals were visualized with the ChemiDocTM Touch imaging system (Bio-Rad, Shanghai).