Brain tissues were homogenized in ice-cold homogenization buffer (4 mM HEPES, pH 7.4; 320 mM sucrose) containing a protease inhibitor mixture (Complete; Roche Diagnostics, IN, USA).
Briefly, brain tissues were homogenized (H) in ice-cold homogenization buffer (10 mM Tris-HCl, pH 7.4, 1 mM EDTA, 1 mM EGTA, 320 mM sucrose (all from Sigma-Aldrich, Rehovot, Israel), 1X protease inhibitor mixture (Sigma-Aldrich Rehovot, Israel or Thermo Scientific, Rockford, USA); and 1X phosphatase inhibitor mixture (Sigma-Aldrich or Thermo Scientific).
Brain tissues were homogenized with Zirconia beads and 700 μl of buffer RA1 including 1% 2-melcaptoethanol by a Beads Homogenizer (Wakenyaku Co., Ltd., Kyoto, Japan), after which 700 μl of 70% ethanol was added to each lysate.
The mouse brain tissues were homogenized in lysis buffer (10 mM Tris-HCL, pH 7.4, 150 mM NaCl, 5 mM EDTA, 0.5% Nonidet P-40, 10 mM Na-β-glycerophosphate, Phosphate inhibitor mixture I and II (Sigma), and complete protease inhibitor mixture (Roche)], using a Diax 900 homogenizer (Sigma).
Brain tissues were homogenized with pre-cooled 1% polyvinylpolypyrrolidone solution (50 mM in pH 7.6 PBS) after isolation.
Brain tissues were homogenized and total RNA was isolated using Trizol (Invitrogen).
100 mg of brain tissues was homogenized in 5× volume of immunoprecipitation buffer (25 mM Tris HCl pH 7.4, 200 mM NaCl, 2 mM EDTA, 0.5 mM EGTA and cocktails of protease and phosphatase inhibitors, (Sigma)).
Total protein from brain tissues was homogenized in lysis buffer containing 50 mM Tris-HCl (pH 7.4), 1 mM phenylmethanesulfonyl fluoride (PMSF), 0.1% leupeptin, and 0.5 M ethylenediaminetetraacetic acid (EDTA) (pH 8.0).
Following a modified version of a previously described protocol [ 30], 100 mg brain tissue were homogenized in 15 mL cold Minimum Essential Medium (MEM) (Gibco, Life Technologies) in a glass tissue grinder (Wheaton), homogenates adjusted to a final concentration of 15% Ficoll and centrifuged at 6000 g and 4°C for 20 min.
Samples from human brain tissue were homogenized in extraction buffer (see blow) by pulsed ultrasonification at 4°C, followed by centrifugation at 10000×g at 4°C for 30 min. The resulting supernatant was used for protein analysis.
Neuronal cells or brain tissue were homogenized in 5 ml of solution A [0.32 M sucrose (34 g/500 ml), 0.5 mM CaCl2 (36 ml/500 ml), 1 mM NaHCO3 (42 mg/500 ml), 1 mM MgCl2 (102 ml/500 ml)] containing protease and phosphatase inhibitors with 12 strokes of a 19×84 mm tissue grinder (Potter Elvehjem, plastic coated) at 800 r.p.m.
