Tris hydrochloride

Glucose, Sucrose, Mannitol, Gelatin, Sorbitol, Sodium chloride, Tris hydrochloride and MgSO₄.7H₂O were purchased from HiMedia Laboratories (chemicals), India. Polyethylene glycol MW6000 (PEG 6000) was supplied by Sigma-Aldrich chemicals. All the chemicals chosen as an excipient for phage lyophilization were previously been proven to maintain the protein viability. All the other chemicals and solvents were obtained at analytic grade from HiMedia Laboratories, India.

Mice represents different housing conditions were euthanized (sodium pentobarbital 45mg/kg; intraperitonially) after the behavioural test. Hippocampus region (n = 6/group) was dissected out from the whole brain, and stored at -70°C. The hippocampus tissue from each individual was used to isolate total RNA, protein and Gas Chromatography-Mass Spectrometry (GC-MS) analysis, and feces (n = 6/group) also collected from the same individuals. The whole brain was dissected out (n = 4/group), and immediately processed for immunohistochemistry.
Hippocampus tissue was homogenized in lysis buffer [Tris-Hydrochloride (50mM; pH 7.5; Cat# RM613; HiMedia, India) Sodium chloride (150mM; Cat# 1.93206.0521; Merck, India), Ethylenediaminetetraactetic acid (EDTA) Disodium salt (50 mM; Cat# RM1195; HiMedia, India), Tergitol (NP-40; 0.1% V/V; Cat# MKCD6607; Sigma-Aldrich, India), Dithiotheritol (DTT,1 mM; Cat# MB070; HiMedia, India) Sodium orthovanadate (Na3Vo4, 0.2Mm; Cat# S6508; Sigma-Aldrich, India), Phenylmethylsulfonyl fluoride (PMSF;0.023mM Cat# P7626; Sigma-Aldrich, India)] following the procedure as reported [40 (link)]. Final supernatant was collected, concentration of each samples were measured using Bio-photometer Plus (Eppendorf Inc., Germany) and stored in aliquots at -80°C.