Mouse monoclonal anti tau antibody tau 5

Rabbit monoclonal anti-Tau s396 phosphorylated antibody, mouse monoclonal anti-tau antibody (TAU-5), rabbit monoclonal anti-PP2Aα Y307 phosphorylated antibody, rabbit monoclonal anti-N-methyl-D-aspartate receptor (NMDAR) 2A antibody, and mouse monoclonal anti-NMDAR2B antibody were acquired from Abcam (Cambridge, GRB). Rabbit anti-PP2Ac subunit antibody was purchased from Neuron Signaling Technology (Massachusetts, USA). Beta-actin mouse monoclonal antibody was provided by ImmunoWay (California, USA). Goat anti-rabbit IgG (H+L)-HRP and goat anti-mouse IgG (H+L)-HRP were provided by Beijing TDY Biotech (Beijing, CHN). BCA protein quantification kit, Coomassie Brilliant Blue staining solution, and ponceau red staining solution were provided by the Beijing Sinoble Biotechnology Center. Protease and phosphatase inhibitors were provided by Roche (Basel, CHE).

An aliquot of 1 × 10⁷ cells of either tau or cotransformant strains as well as the WT were pelleted, supernatant removed and the pellet resuspended in 75 μL of Laemmli 2× sample buffer (4% SDS, 20% glycerol, 0.004% bromophenol blue and 0.125 M Tris-Cl, pH 6.8). Samples were boiled for 10 min and 15 μL loaded to a 10–15% SDS-PAGE gel and transferred to a PVDF membrane (Bio-Rad, Hercules, CA, United States). To detect tau, membranes were incubated with mouse monoclonal anti-tau antibody [Tau-5] (Abcam, Cambridge, United Kingdom, ab80579) (1:2,000), to detect α-synuclein membranes were incubated with mouse monoclonal anti-alpha-synuclein antibody [syn211] (Abcam, Cambridge, United Kingdom, ab80627) (1:2,000) and to detect phosphorylated tau membranes were incubated with rabbit monoclonal recombinant anti-tau (phosphor S404) antibody [EPR2605] (Abcam, Cambridge, United Kingdom, ab92676) (1:500). This was followed by incubation with (goat) anti-mouse or anti-rabbit Cross-Adsorbed Secondary antibody Alexa-Fluor^TM 647 (Thermo Fisher Scientific, Waltham, MA, United States, A32728) (1:1,000). Proteins were visualised using the STORM image analyser.