Recombinant human alpl

The following were purchased from ThermoFisher Scientific (Carlsbad, CA, USA): MEMα with or without phenol red; MEMα with or without ascorbic acid (vitamin C); MEMα with or without calcium (Ca 2+ ); penicillin/streptomycin; L-glutamine; TaqMan gene expression assays for ALPG (Hs00741068_g1), ALPI (Hs00357579_g1), ALPL (Hs10129144_m1), ALPP (Hs00740632_gH), GAPDH (Hs99999905_m1); universal PCR master mix. Recombinant human ALPL was purchased from R&D Systems (Minneapolis, MN, USA). Rabbit anti-human ALPL antibody, calf intestinal ALP (CIP, ≥10 DEA U/mg), disodium β-glycerophosphate (βGP), phospho(enol)pyruvate monosodium (PEP), pamidronate, dexamethasone, and Alizarin Red S solution were purchased from Sigma-Aldrich (St. Louis, MO, USA). CIP (≥10,000 U/mL), shrimp hepatopancreas ALP (≥1000 unit/mL), and p-nitrophenyl phosphate kits were purchased from New England BioLabs (Ipswich, MA, USA). FBS was purchased from Atlanta Biologicals (Flowery Branch, GA, USA). Disodium α-glycerophosphate hydrate (1 M of αGP; Glycophos) was purchased from Fresenius Kabi (Lake Zurich, IL, USA), and glycerophosphoric acid (NSC 9231) was obtained from the Developmental Therapeutics Program (NCI, USA). Collagen Type I, rat tail, stock solution was obtained from BD Biosciences Discovery Labware (Waltham, MA, USA).