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2 protocols using 4 methylmorpholine nmm

1

Optimizing Peptide Synthesis and Cell Culture Techniques

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Fmoc-protected amino acids were purchased from AAPPTEC (Louisville, KY) and EMD Biosciences (San Diego, CA). Resins were obtained from Chem-Implex (Wood Dale, IL), N-methyl-2-pyrrolidinone (NMP) from Advanced ChemTech (Louisville, KY), and 4-methylmorpholine (NMM) from Sigma-Aldrich (St. Louis, MO); all were used without further purification. Acetic anhydride (Ac2O) was purchased from Fisher Scientific (Pittsburgh, PA). All the cancer cell lines and media were obtained from American Type Culture Collection (ATCC, Manassas, VA). PathHunter assay kit was from DiscoverX technologies (Fremont, CA). For Western blot experiment, Novex® 4–20% tris-glycine gels and cell lysis buffer were ordered from Life Technologies (Grand Island, NY) and antibodies from Abcam, Inc. (Cambridge, MA). The antibodies for the housekeeping protein glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX).
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2

Synthesis of Hyaluronic Acid Conjugates

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Sodium salt Hyaluronate (HA) isolated from Streptococcus equi with average Mw 1.2 MDa was purchased from Caldic (Origgio, Varese, Italy). Ornithine methyl ester (H-Orn-OMe. 2HCl) was purchased from Bachem. 4-methylmorpholine (NMM), NaCl, 0.025 M sodium tetraborate, sulfuric acid, and acetonitrile were purchased from Sigma–Aldrich. 2-chloro-dimethoxy-1,3,5-triazine (CDMT) purchased from TCI. The phosphate buffer saline (PBS) was purchased from Sigma–Aldrich (Sigma-Aldrich SRL, Milano, Italy).
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