2 amino 2 methyl propan 1 ol

All the aqueous solutions were prepared with Millipore DirectQ ultra pure apyrogenic water. Bovine serum albumin (BSA), trichloroacetic acid (TCA), tris-hydroxymethyl-amino-methane (Tris), N-(2-hydroxyethyl)piperazine-N’-ethanesulfonic acid (HEPES), 2-amino-2-methyl-propan-1-ol (AMPOL), sodium dodecylsulfate (SDS), p-nitrophenyl phosphate disodium salt (pNPP), dexamethasone, β-glycerophosphate, polyoxyethylene-9-lauryl ether (polidocanol), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS), Rhodamine 6G, collagens type I (from calf skin), type II (from bovine nasal septum), types I+III (from horse tendon) and collagenase were obtained from Sigma-Aldrich (St Louis, MO, USA). Sodium phosphate, magnesium chloride, acetic acid and chloroform were acquired from Merck (São Paulo, SP, Brazil). Plastic culture flasks (75 cm²) were purchased from Corning (Cambridge, MA, USA). 96 well plates, α-MEM, fetal bovine serum, ascorbic acid, gentamicin, penicillin-streptomycin and fungizone were supplied by Gibco-Life Technologies (Grand Island, NY, USA). Collagen II monoclonal antibody (2B1.5, MA5-12789) and Goat anti-mouse Alexa Fluor 488 (A11001) were purchased from Invitrogen (Massachusetts, USA). Analytical grade reagents were used without further purification.

All the solutions were prepared by using dust-free Milli-Q water (γ= 72.8 mN m-1 and resistivity of 18.2 MΩ cm). Sodium dodecylsulfate (SDS), polyoxyethylene-9-lauryl ether (polidocanol, C12(EO)9), potassium bromide, magnesium chloride, sodium chloride, p-nitrophenylphosphate (pNPP), chloroform, 2-amino-2-(hydroxymethyl)propane-1,3-diolhydrochloride (Tris), and 2-amino-2-methyl-propan-1-ol (AMPOL) were purchased from Sigma Chemical Co. [(2R)-2,3-Di(hexadecanoyloxy)propyl] 2-(trimethylazaniumyl)ethyl phosphate (dipalmitoylphosphatidyl-choline, DPPC) (99+%) and (8S,9S,10R,13R,14S,17R)-10,13-dimethyl-17-[(2R)-6-methylheptan-2-yl] 1,2,6,7,8,9,11,12,14,15,16,17-dodecahydrocyclopenta[a]phenanthren-3-one (cholestenone, A-Chol) (99+%) were obtained from AVANTI Polar Lipids. Methanol (MeOH, 99+%) was supplied by J.T. Baker. 3-N,3-N,6-N,6-N-tetramethylacridine-3,6-diamine (acridine orange, AO) was acquired from Acros Organic. TNAP was obtained from osteoblasts primary cultures as previously reported elsewhere [15 (link)]. Calbiosorb was purchased from Merck-Millipore.