Mc3t3 e1 subclone 4 atcc crl 2593

Doxorubicin∙HCl (>99 %) was purchased from TSZ Chem BIOTANG Inc. AzideFluor 488 (HPLC), 5-bromovaleric acid, and levulinic acid were purchased from Sigma Aldrich. Rink Amide MBHA resin LL (100–200 mesh, 0.52 meq) was purchased from Novabiochem. Aminooxy-5(6)-TAMRA was purchased from Biotium, Inc. MC3T3-E1 Subclone 4 (ATCC® CRL-2593™) was purchased from ATCC. Fmoc-protected amino acids and O-(1H-6-Chlorobenzotriazole-1-yl)-1, 1, 3, 3-tetramethyluronium hexafluorophosphate (HCTU) were obtained from Peptides International.

All the chemicals were obtained from Sigma Aldrich (St. Louis, MO) and cell culture reagents were purchased from Gibco (Carlsbad, CA), unless otherwise stated. Murine cell line, MC3T3-E1 subclone 4 (ATCC^® CRL-2593™), which is competent to produce mineralizing bone-like matrix was purchased from American Type Culture Collection. These cells were grown at 1 ×10⁵ cells /100mm plate in growth medium [αMEM (HyClone, Logan, UT) without ascorbic acid, 10% FBS (HyClone, Logan, UT), 1% penicillin/streptomycin, and 2 mM glutamine] (Wang et al., 1999 ). Cells were subcultured when they reached 50–60% confluence, usually after 3–4 days after plating. All the experiments were done in triplicate with cells of passage 8–12, at 37° C, under 5% CO₂ and in humidified conditions.