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4 protocols using n boc ethylenediamine

1

Synthesis and Characterization of Tartrate Derivatives

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All reagents and solvents were purchased from Sigma-Aldrich (Milwaukee, WI) and used as received unless otherwise mentioned. Di-tert-butyl L-tartrate22 (link) and di-2-bocaminoethyltartramide23 were prepared as previously published. Anhydrous dimethylformamide (DMF) was dried over 4 Å molecular sieves at room temperature at least overnight prior to use. N-Boc-ethylenediamine was purchased from Alfa Aesar (Ward Hill, MA). 1-(3-dimethylaminopropy-l)-2-ethylcarbodiimide hydrochloride (EDC·HCl) was purchased from AK Scientific (Union City, CA). Silicon wafers were purchased from Ted Pella, Inc. (Redding, CA). For cell experiments, reagents include human buffy coats purchased from the New York Blood Center (Long Island City, NY), penicillin/streptomycin purchased from Lonza (Basel, Switzerland), Dulbecco’s modified eagle medium (DMEM) and Vybrant® MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell proliferation assay kit purchased from ThermoFisher Scientific (Waltham, MA).
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2

Chondrocyte Differentiation Protocol

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Collagenase type V, exendin-4 (Ex4), manganese (II) chloride (MnCl2·4H2O, 99%), iron (III) acetylacetonate (Fe(acac)3, 99.9%), methyl poly(ethylene glycol) (mPEG, M.W. = 2000), N-hydroxysuccinimide (NHS), N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide (EDC), oleic acid (90%), oleylamine (90%), osmium tetroxide (1%) and Prussian blue were from Sigma–Aldrich (St. Louis, MO, USA). Acryloyl chloride (96%) and N-Boc-ethylenediamine (98%) were from Alfa Aesar (Ward Hill, MA, USA). Benzyl ether and (3-aminopropyl) triethoxy silane (APTES, 98%) were from Fluka (Buchs, SG, Switzerland). N-Hydroxybenzotriazole (HOBt) and (benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate (PyBOP) were from NovaBiochem (Beeston, NTH, UK). RPMI-1640 medium was from GIBCO BRL (Grand Island, NY, USA). Polyethylene (PE-50) tubing was from Clay Adams (Parsippany, NJ, USA). Guinea pig anti-swine insulin antibody was from Dako (Carpinteria, CA, USA). Rabbit polyclonal anti-SOX9 antibody (E-9) was from EMD Millipore Corporation (Temecura, CA, USA).
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3

Synthesis and Characterization of Multifunctional Nanoprobes

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Iron(III) acetylacetonate ([Fe(acac)3], 99.9%), manganese(II) chloride (MnCl2•4H2O, 99%), methyl poly(ethylene glycol) (mPEG, M.W. = 2,000), oleic acid (90%), oleylamine (90%), N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC), IR-783, fluorescein isothiocyanate (FITC), Prussian blue, and nuclear red fast were purchased from Sigma-Aldrich (St. Louis, MO, USA). Acryloyl chloride (96%) and N-Boc-ethylenediamine (98%) were purchased from Alfa Aesar (Ward Hill, MA, USA). (3-Aminopropyl) triethoxy silane (APTES, 98%) and benzyl ether were purchased from Fluka (Buchs, SG, Switzerland). (Benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate (PyBOP) and N-hydroxybenzotriazole (HOBt) were purchased from NovaBiochem (Beeston, NTH, UK). BCA (bicinchoninic acid) protein assay reagent kit was purchased from Pierce (Rockford, IL, USA). Molecular-porous membrane tubing (M.W. = 12~14 kDa and M.W. = 50 kDa) was purchased from Spectrum (Houston, TX, USA). The primary antibodies (mouse anti-EGFR and anti-β-actin) and secondary antibody (Rabbit polyclonal antibody to mouse IgG-H&L) were purchased from Abcam (Cambridge, MA, USA), and all the chemicals were used directly without any further purification unless otherwise stated.
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4

Nanomaterial Synthesis and Characterization

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Collagenase type XI, exendin-4 (Ex4), Histopaque®-1077, manganese (II) chloride (MnCl2·4H2O, 99%), iron (III) acetylacetonate (Fe(acac)3, 99.9%), methyl poly(ethylene glycol) (mPEG, M.W. = 2000), N-hydroxysuccinimide (NHS), N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide (EDC), oleic acid (90%), oleylamine (90%), osmium tetroxide (1%), propidium iodide (PI) and Prussian blue were purchased from Sigma–Aldrich (St. Louis, MO, USA). N-Boc-ethylenediamine (98%) and Acryloyl chloride (96%) were from Alfa Aesar (Ward Hill, MA, USA). Benzyl ether and (3-Aminopropyl) triethoxy silane (APTES, 98%) were from Fluka (Buchs, Switzerland). N-hydroxybenzotriazole (HOBt) and (Benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate (PyBOP) were from NovaBiochem (Beeston, UK). RPMI-1640 medium and Dulbecco’s modified Eagle’s medium (DMEM) were from GIBCO BRL (Grand Island, NY, USA). Insulin radioimmunoassay (RIA) kit was from Millipore Corporation (Billerica, MA, USA). Polyethylene (PE-50) tubing was from Clay Adams (Parsippany, NJ, USA). Guinea pig anti-swine insulin antibody and rabbit anti-human glucagon antibody were from Dako (Carpinteria, CA, USA). Guinea pig anti-human insulin antibody (ab7842) was from Abcam (Cambridge, MA, USA).
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