Rabbit anti glua1

Primary antibodies included: mouse anti-GluA1 (NeuroMAB; 1:10 IHC), rabbit anti-GluA1 (Cell Signaling Technologies; 13185, 1:1,000), mouse anti-GluA2 (NeuroMAB; 1:10 IHC), mouse anti-GluA2 (MAB1189; 1:500), chicken anti-Homer (SySy; 160006, 1:500), mouse anti-SynGAP (Thermo Fisher Scientific; PA1-046, 1:500 IHC, 1:5,000 WB), mouse anti-PSD95 (Thermo Fisher; MA1-045, 1:1,000 WB), rabbit anti-Synaptophysin (PA1-1043, 1:5,000 WB), mouse anti-Cdk5 (Invitrogen; AHZ0492, 1:500), mouse anti-WAVE1 (mAb K91/361; NeuroMab), goat anti-PPA2c (Santa Cruz; sc-6110), rabbit anti-CaMKIIα (Abcam; EP1829Y, 1:1,000), rabbit-anti-p-Synapsin1-Ser 551 (Abcam; Ab32532), rabbit anti-p35 (Cell Signaling; 2680, 1:250), mouse anti-GAPDH (Millipore; MAB374, 1:4,000), and Phalloidin Alexa 647 (Thermo Fisher Scientific; A22287, 1:200). Secondary antibodies included: donkey anti-mouse Alexa 488 (Thermo Fisher Scientific; R37114, 1:200), donkey anti-mouse Dy 488 (Abcam; 96875, 1:500), donkey anti-chicken Alexa 488 (Jackson ImmunoResearch; 703-545-155), and LI-COR (anti-mouse, anti-goat, and anti-rabbit IRDye 680 and 800; 1:10,000).

Activity-dependent responses in hippocampal synaptosomes were analyzed by FASS-LTP, as previously described^{26 (link),27 (link)}. FASS-LTP consists of chemical LTP (cLTP) stimulation directly in crude P2 synaptosomal fractions, immunofluorescence labeling for surface GluA1 and neurexin-1β, and flow cytometry analysis. Briefly, fresh crude synaptosome P2 fractions were obtained from the hippocampi of mice injected with AAVs and stimulated with glycine (500 μM) and KCl (50 mM). For surface immunolabeling, primary antibodies were rabbit anti-GluA1 (Cell Signaling #13185; 1:400) and mouse anti-Nrx1β (UC Davis/NIH NeuroMab Facility, 75–216; 2.5 μg/ml). Secondary antibodies were anti-rabbit-Alexa-405, anti-rabbit-Alexa-488 and anti-mouse-Alexa-647 antibodies (Life Sciences), at 2.5 μl/ml. Samples were protected from light, maintained at 4 °C and immediately run on a flow cytometer (Novocyte, ACEA Biosciences, Inc); 20,000 events were collected and analyzed for each sample with an event rate of approximately 500/sec. Analysis was performed using the FlowJo v10.3 software (LLC).

The primary antibodies used were: mouse anti-tubulin (1:30,000, #T9026, Sigma), rabbit anti-GAPDH (1:5,000, #sc-25778, Santa Cruz), rabbit anti-GluN2A (1:1,000, #M264, Sigma), rabbit anti-GluN2B (1:1,000, #718600, Invitrogen), mouse anti-GluN2D (1:1,000, #MAB5578, Millipore), rabbit anti-GluA1 (1:1,000, #13185, Cell Signaling), mouse anti-GluA2 (1:1,000, #75–002, Neuromab), mouse anti-GluA3 (1:1,000, #MAB5416, Millipore), polyclonal anti-Rph3A (1:2,000, Protein Tech, #11396-1-AP), mouse anti-PSD-95 (1:1,000, #K28/43, Neuromab), rabbit anti-tyrosine hydroxylase (1:10,000, #AB152, Millipore), rabbit anti-phospho-extracellular signal-regulated kinase (ERK) 44/42 (1:1,000, Cell Signaling, #9101), rabbit anti-ERK 44/42 (1:1,000, #9102, Cell Signaling), rabbit anti-phospho-cAMP responsive element binding protein (CREB) (1:1,000, #9198, Cell Signaling), and rabbit anti-CREB (1:1,000, #9197, Cell Signaling). The secondary horseradish peroxidase-linked antibodies used were goat anti-rabbit (1:10,000, #172-1019, Biorad) and goat anti-mouse (1:10,000, #172-1011, Biorad).