Rabbit anti glua1 pab

The following primary antibodies were used in this study: purified rabbit anti-ZDHHC8 pAb (1:200; Abcam); rabbit anti-ZDHHC8 pAb (1:100; Bioss); rabbit anti-ZDHHC5 pAb (1:500; Proteintech Group, Inc.); rabbit anti-GADPH mAb (1:1000; Proteintech Group, Inc.); guinea pig anti-MAP2 mAb (1:200; Synaptic Systems); mouse anti-GFAP mAb (1:100; Proteintech Group, Inc.); chicken anti-GAD67 mAb (1:300; Synaptic Systems); mouse anti-PSD95 mAb (1:50; Proteintech Group, Inc.); guinea pig anti-VGLUT1 mAb (1:200; Synaptic Systems); mouse anti-gephyrin mAb (1:50; Santa Cruz Biotechnology); DAPI (1:50; Sigma-Aldrich); rabbit anti-GluA1 pAb (1:500; Abcam); rabbit anti-GluA2 pAb (1:200; Proteintech Group, Inc.); rabbit anti-GluA3 pAb (1:500; Bioworld Technology, Inc.); rabbit anti-GluA4 pAb (1:200; Proteintech Group, Inc.); Anti-Sodium Potassium ATPase mAb (1:1000; Abcam); and GABAAR β2/3 (1:200; Proteintech Group, Inc.).

Mice were anesthetized and decapitated, and brain tissue was quickly extracted. The hippocampus was rapidly dissected on ice, incubated in modified RIPA buffer (Beyotime, Nantong, China) on ice for 15 min, and centrifuged at ~200,000 × g for 10 min at 4 °C. The supernatant was collected and supplemented with 100 µl protein A/G-agarose beads overnight at 4 °C on a wheel. The beads were then allowed to sediment at the bottom of the tube, and the supernatant was collected. ZDHHC8 (1:200, Abcam), rabbit anti-GluA1 pAb (1:500; Abcam), rabbit anti-GluA2 pAb (1:200; Proteintech Group, Inc.), rabbit anti-GluA3 pAb (1:500; Bioworld Technology, Inc.), and rabbit anti-GluA4 pAb (1:200; Proteintech Group, Inc.) antibodies were incubated in equal amounts with the supernatant. Protein A/G-agarose beads were added to the supernatant, and the supernatant was incubated for 2 h at room temperature on a wheel. The beads were then washed three times with RIPA buffer and submitted to SDS-PAGE. The mixture was boiled for 10 min. The Western blots were detected using ZDHHC8 and GluA1, GluA2, GluA3, and GluA4 antibodies for immunoprecipitation^{21 (link)}.