Sheep anti trem2

The following antibodies were used in this study: Mouse anti-β-Amyloid (BioLegend, SIG-39320), rabbit anti-β-Amyloid (Proteintech Group, 25524-1-AP), mouse anti-Galectin-3 (BioLegend, 126702), goat anti-Galectin-3 (R&D Systems, AF1154), goat anti-GPNMB (R&D Systems, AF2330), mouse anti-GAPDH (Proteintech Group, 60004-1-Ig), rat anti-mouse LAMP1 (BD Biosciences, 553792), LAMP2 (Developmental Studies Hybridoma Bank, GL2A7-c), goat anti-CathB (R&D Systems, AF965), goat anti-CathD (R&D Systems, AF1029), goat anti-CathL (R&D Systems, AF1515), sheep anti-TREM2 (R&D Systems, AF1729), rabbit anti IBA-1 (Wako, 01919741), goat anti-AIF-1/Iba1 (Novus Biologicals, NB100-1028), rat anti-CD68
(Bio-Rad, MCA1957), rabbit anti-TFE3 (Sigma, HPA023881), and sheep anti-PGRN (R&D Systems, AF2557). PS1-related peptides were detected using Ab14, a rabbit polyclonal antibody generated against amino acids 1-25 of PS1. Rabbit anti-BACE1 antibody were raised against the sequence CLRQQHDDFADDISLLK from 485 to 501 of BACE1 protein (Yan et al., 2001) .
The following reagents were also used in the study:

Free-floating 30 μm sections were washed once in PBS for 10 minutes. The sections then underwent antigen retrieval in SSC buffer (10mM, pH 9.0) for 30 minutes at 80°C. The tissues were permeabilised by washing three times for 10 minutes in 0.3% Triton X-100 in PBS (PBST). Non-specific binding was blocked for 1 hour in 10% donkey serum, followed by incubation with the appropriate primary antibodies in blocking solution at 4°C overnight (1:1000 rabbit anti-IBA1 (FujiFilm Wako Chemicals, Cat # 019-19741) & 1:500 sheep anti-TREM2 (R&D Systems, Cat # AF1729)). The next day sections were washed three times for 10 minutes in PBST followed by a 2-hour incubation in the dark with secondary antibodies in blocking solution (1:1000 donkey anti-rabbit Alexa Fluor 594 (ThermoFisher Scientific, Cat # A32 and 1:1000 donkey antisheep Alexa Fluor 647 (Abcam, Cat # AB150179)). Finally, nuclei were counterstained with 4ʹ,6-diamidino-2phenylindole (DAPI) and plaques labelled with Amytracker520 (Ebba Biotech, Sweden).