Anti rabbit txrd

ThioS stain was done as previously described 62 (link). Briefly, slices were washed twice for 10 min in PBS. Next, slices were put in a blocking solution (TBS, 0.3% TritonX-100, 3% goat serum) for 2 h. Then slices were put in anti-NeuN (Millipore, ABN78) 1:1000 in TBS with 0.3% Triton X-100 (TBS-T) overnight. The next day, slices were washed three times in TBS-T, then put in anti-rabbit-TXRD (Southern Biotech, 4010-07) 1:300 in TBS-T overnight. The next day, 3 TBS-T washes were performed, then the slices were stained in 1% ThioS (Sigma, T1892) for 9 min. To de-stain, slices were washed twice with water, once with 70% ethanol, then twice with water. Slices were finally washed again in TBS then mounted onto Superfrost Plus slides (VWR, 48311-703). AF1+DAPI (EMS, 17970-125) mounting media was added, then the coverslip. Slides were imaged using a Zeiss Axio Imager.M2 the next day.

Brain slices were mounted onto Superfrost Plus slides (VWR, 48311-703) (62, 63). Next, the slides were put in 4% PFA for 4 min, 2 TBS washes, then 70% formic acid (Sigma, F0507) for 2 min. Next, slides were washed twice with TBS, then put in TBS with 0.1% Triton-X for 15 min. After, slides were blocked in TBS-B (TBS, 0.1% Triton-X, 2% BSA) for 30 min. Anti-6E10, a Aβ 1-16 antibody, (Biolegend, 803015) at 1:1000 and anti-NeuN (Millipore, ABN78) 1:1000 were added in TBS-B for two days at room temperature. After washing in TBS, anti-mouse-alexa-488 (Southern Biotech, 1010-30) at 1:500 and anti-rabbit-TXRD (Southern Biotech, 4010-07) at 1:300 were added for 6 h, or up to overnight. After secondary, slides were washed in TBS, then AF1+DAPI (EMS, 17970-125) mounting media was added, then the coverslip. Slides were imaged using a Zeiss Axio Imager.M2 the next day.