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3 protocols using 4 4 6 dimethoxy 1 3 5 triazin 2 yl 4 methyl morpholinium chloride dmt mm

1

EGCG Gelatin Synthesis Protocol

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EGCG was purchased from Bio Verde Inc. (Kyoto, Japan), N-methylmorpholine (NMM) was purchased from Nacalai Tesque Inc. (Kyoto, Japan), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMT-MM) was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan), and type A gelatin from porcine skin was acquired from Sigma-Aldrich (St. Louis, MO, USA).
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2

Hyaluronic Acid-Based Bioactive Hydrogel

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Hyaluronic acid (HA) (1 MDa, Humedix, Anyang, Korea), methyltetrazine-PEG4-amine hydrochloride salt (Click Chemistry Tools, AZ, USA), trans-cyclooctene-amine hydrochloride salt (Click Chemistry Tools, AZ, USA) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM) (Tokyo chemical industry, Tokyo, Japan) and Substance P (SP; RPKPQQFFGLM) (GenScript, NJ, USA) were used as received. Substance P analog (SPa; RISPQQRDALA), BMP-2 mimetic peptide (BP; KIPKASSVPTELSAISTLYL), fluorescein isothiocyanate-conjugated SPa (SPa-F), fluorescein isothiocyanate-conjugated SP (SP–F), fluorescein isothiocyanate-conjugated BP (BP–F) were purchased from Biostem (Suwon, Korea). 1,4-[D]iaminobutane (Sigma, MO, USA), protamine, sodium azide, propargyl amine, IR-783, indocyanine green (ICG) and PKH26 red fluorescent cell linker kit were purchased from Sigma (MO, USA). Low-glucose Dulbecco's modified Eagle's medium (DMEM), Dulbecco's phosphate-buffered saline (DPBS), penicillin-streptomycin (PS), 0.05 % trypsin-EDTA and fetal bovine serum (FBS) were purchased from Gibco BRL (Carlsbad, CA, USA).
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3

Enantioselective Derivatization and Quantification

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The enantiomers (sodium D-LA and L-LA) and a racemic mixture of sodium LA was purchased from Sigma Aldrich (St. Louis, MO, USA). Methanol (MeOH) and ethanol (EtOH) of HPLC grade, propionitrile (PrCN) and trifluoroacetic acid (TFA) were purchased from FUJIFILM Wako Pure Chemical Industries (Osaka, Japan). Acetonitrile (MeCN) of HPLC grade was obtained from Nacalai Tesque (Kyoto, Japan). 4-Nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) were products of Tokyo Chemical Industry (Tokyo, Japan). Water was purified by a Milli-Q Integral 3 system (Merck, Darmstadt, Germany). All other reagents were of the highest reagent grade and were used without further purification.
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