Methanol meoh

Trypsin, sodium dihydrogen phosphate monohydrate, di-sodium hydrogen phosphate dihydrate, 1,4-dithioerythritol (DTE), 2-iodoacetamide (IAA), sodium borohydride, Dowex 50WX8 ion exchange resins, sodium hydroxide (NaOH), iodomethane, sDHB (9:1 mixture of 2,5-dihydroxybenzoic acid and 2-hydroxy-5-methoxy benzoic acid) and dextran were purchased from Merck (Taufkirchen, Germany). Acetonitrile (ACN) and anhydrous dimethyl sulfoxide (DMSO) were purchased from VWR International GmbH (Dresden, Germany). Trifluoroacetic acid (TFA) and acetic acid were purchased from Thermo Fisher Scientific (Schwerte, Germany). Peptide N-glycosidase F (PNGase F) was purchased from N-Zyme Scientifics (Doylestown, PA, USA). Methanol (MeOH) was purchased from Carl Roth GmbH + Co. KG (Karlsruhe, Germany). Milli-Q grade water was produced using a MilliQ Plus water purification system (Merck KGaA, Darmstadt, Germany). All the reagents were obtained in p.a. quality.

A freeze-dried biomass of Thermoplasma acidophilum (TransMIT GmbH, Development of colloidal drug formulations, Giessen, Germany) was stored at −20 °C until usage. Phospholipon 100H (Ph; commercial phosphatidylcholine including stearic and palmitic acid) and 5(6)-carboxyfluorescein (CF) were purchased from Lipoid GmbH (Ludwigshafen, Germany) and Fluka, (Buchs, Switzerland), respectively. Fetal calf serum (FCS) was obtained from PAA (Coelbe, Germany), Triton X-100 was provided by Carl Roth (Karlsruhe, Germany). The lung surfactant Alveofact was provided by Lyomark Pharma (Oberhaching, Germany). Chloroform (CHCl₃) and methanol (MeOH) were both purchased from Carl Roth (Karlsruhe, Germany) and the Chloroform:Methanol (CHCl₃:MeOH) (2:1 v/v) mixture was stored at room temperature until it was used. HEPES buffer solution (HBS) was prepared with 40 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), 5 mM Glucose (Dextrose), and 120 mM NaCl and adjusted to pH 7.4. Phosphate-buffered saline (pH 7.4) composed of 1.3 mM KH₂PO₄, 7.4 mM Na₂HPO₄ × 2 H₂O, 129 mM NaCl, and 2.5 mM KCl, which were dissolved in bidistilled water. All chemicals and solvents used in the experiments were of analytical grade.