At8 anti p tau pser202 thr205

Equal amounts of protein from (30 μg, mouse cortex was used) were separated by electrophoresis in precast 4-12% Bis-Tris Gels (Bio-Rad) and transferred to nitrocellulose membranes. The membranes were hybridized with the following primary antibodies as indicated: TYROBP (1:500, antibody provided in collaboration with Dr Richard W. Cho at Cell Signaling Technology); AT8 anti-pTAU pSer202/Thr205 (1:1000, Thermo Scientific, #MN1020); anti-GAPDH (1:2000, Santa Cruz, #SC-32233), anti-p-TAU Thr205 (1:1000, Invitrogen, #44-738G), anti-TAU HT7 (1:1000, Invitrogen, #MN1000). Secondary antibodies included: peroxidase-labeled anti-mouse IgG (1:2000, Vector Laboratories); peroxidase-labeled anti-rabbit IgG (1:2000, Vector Laboratories); peroxidase-labeled anti-rat IgG (1:2000, Vector Laboratories). ECL (Pierce, #32106) was used to reveal the immunoreactive proteins, and images were acquired using a Fujifilm ImageReader LAS-4000. Membranes were stripped using a stripping buffer (Thermo Scientific, #46430) when required. Luminescent immunoreactive protein bands were quantified using Fiji software (ImageJ).