Mab1560

Western blot analysis and densitometry were carried out by using standard procedures, according to Borreca et al. (2018) (link).
The following antibodies were used: anti-Aβ/APP 6E10 (4–9aa) mouse MAB1560 Chemicon; anti-Alzheimer precursor protein 22C11 (66–81aa of N-terminus) mouse APP-MAB348 Chemicon Temecula-CA; anti-pan tau protein H150 (1–150aa of N-terminus) rabbit sc-5587 Santa Cruz Biotechnology; anti-pan tau protein DC25(microtubule binding repeat) mouse T8201 Sigma-Aldrich; tau 21 (21–36aa of N-terminus) rabbit AHB0371 Biosource International (USA); anti-N-tau (45–73aa) DC39N1 mouse T8451 Sigma-Aldrich; neuronal marker β III tubulin antibody mouse ab78078 (clone 2G10) Abcam; GAPDH antibody (6C5) mouse sc-32233 Santa Cruz Biotechnology; activity-regulated cytoskeleton-associated protein (C-7) mouse sc-17839 Santa Cruz Biotechnology; glial fibrillary acidic protein antibody rabbit Z0334 Dako; Iba1 antibody rabbit Wako 016-20001 (for WB).

Mice were deeply anesthetized and transcardially perfused with ice‐cold 0.9% saline, followed by 4% paraformaldehyde dissolved in 0.1 M phosphate‐buffered saline (PBS). The fixed, cryoprotected mouse brains were frozen and then cut into 35‐μm‐thick coronal sections. Free‐floating sections were washed three times in PBS, following which they were incubated for 1 hr in 5% normal goat serum in PBS containing 0.3% Triton X‐100. Sections were incubated overnight at 4°C with the following primary antibodies: 6E10 mouse monoclonal anti‐Aβ (1:1,000, Chemicon; MAB1560), rat monoclonal anti‐bromodeoxyuridine (BrdU; 1:100, AbD Serotec; OBT0030), goat anti‐doublecortin (DCX; 1:200, Santa Cruz Biotechnology; SC8066), mouse anti‐glial fibrillary acidic protein (GFAP; 1:200, Abcam; AB4648), and mouse monoclonal anti‐BDNF (1:500, Abcam; ab203573) antibody. After washing three times in PBS, the sections were incubated with the corresponding Invitrogen Alexa Fluor secondary antibodies in PBS for 1 hr at room temperature. The sections were then washed and mounted, and coverslips were secured on the slides using a mounting medium (Vector Laboratories, Inc. Burlingame, CA; H1200). Confocal images were captured using a Nikon confocal Axiovert microscope (Carl Zeiss, Göttingen, Germany; LSM510).