Pbs tablet

The analytical standard of OTA was supplied by Sigma-Aldrich (St. Louis, MO, USA). All solvents used for the preparation of the mobile phase were HPLC grade and obtained from Merck (Darmstadt, Germany). Methanol and hexane used for extraction were of analytical grade supplied by Sigma-Aldrich. All homogenized mixtures and eluates were filtered through Whatman no. 4 and 0.45 mm membrane filters respectively (Whatman plc, Maidstone, UK). De-ionized water was obtained with a Millipore Elix Essential purification system (Bedford, MA, USA). OCHRA PREP immunoaffinity columns were supplied by R-Biopharm, Rhone limited, and used for SPE and cleanup. These columns have a concentration capacity of 100 ng/mL with at least 90% recovery. Phosphate-buffered saline (PBS) was prepared by dissolving PBS tablets (Sigma-Aldrich) in distilled water. Sodium chloride (≥ 99.0%) was sourced from Sigma-Aldrich. Six-point calibration was made using the pure Ochratoxin A standard at concentrations of 5 µg/kg, 10 µg/kg, 15 µg/kg, 20 µg/kg, 25 µg/kg and 30 µg/kg. Linearity was accepted at 0.99 or 99% for the regression curve (CEN official method EN14123³³ ).

Twenty-four h after the last dose of FLX or vehicle, Mecp2 HET and WT mice, were anaesthetized with 75 mg/kg ketamine plus 1 mg/kg medetomidine and perfused through the heart using a peristaltic pump (Mityflex, ANKO, Bradenton, FL) with Ice-cold 0.01 M phosphate buffer-saline (PBS Tablets; Sigma, Cat. # 5564) at 10 mL/min for 3 min followed by 4% para-formaldehyde (PAF) solution in PBS for 5 min. The brain was removed and post-fixed in 4% PAF for 4 h, at 4 °C. Brains were then dipped in a 20% sucrose solution and stored at 4 °C overnight. Brains were removed from the sucrose solution, damped with Kleenex, frozen in n-pentane at − 45 °C for 3 min and stored at − 80 °C until sectioning.
Brain sections (25 μm) for IHC were cut with a cryostat (CM1850-UV, Leica Microsystems, Buccinasco, Italy) at − 17 °C and mounted on gelatin-coated slides. Consecutive brain slices containing the areas of interest were mounted onto glass slides for 3,3′-diaminobenzidine tetrahydrochloride (DAB) staining.

Na₂HPO₄, NaH₂PO₄, KCl, HCl, NaOH, and PBS tablets were purchased from Sigma Aldrich (St. Louis, MO, USA). PBS tablets were dissolved in 200 mL deionized water. Chloroform (CHCl₃), tetrahydrofuran (THF), and absolute ethanol (EtOH) were bought from Merck KGaA (Darmstadt, Germany). The polymers monocarboxy terminated poly(N-isopropyl acrylamide) (PNIPAAM-COOH), poly(acrylic acid) (PAA), and the adhesion promoter poly(glycidyl methacrylate) (PGMA) were purchased and characterized from Polymer Source, Inc. (Dorval, QC, Canada). Here, PGMA with molecular weight (MW) of 17.500 g/mol was chosen, whereas PAA had a MW of 26.500 g/mol and PNIPAAM of 47.600 g/mol. The positive photoresist AZ5214e was obtained by MicroChemicals GmbH, (Ulm, Germany). Cell culture reagents such as McCoy’s 5A, FBS, trypsin, L-glutamine, pen/strep, and sterile water were obtained from Biochrome AG (Berlin, Germany). Phosphate buffer solutions (0.1 M; Na₂HPO₄/NaH₂PO₄) with pH ranges from 5.7 to 8.0 were prepared.
Saos-2 cells were donated by Prof. Wiesmann, TU Dresden, IfWW.