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3 protocols using 1 4 7 10 tetraazacyclododecane 1 4 7 10 tetraacetic acid dota

1

Trace Metal Purification Protocol

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Optima grade HCl comes from Aristar Ultra, VWR, West Chester, PA. Natural calcium (natCa, 99.99%) in dendritic chunks comes from Sigma-Aldrich, St. Louis MO. UTEVA (100–150 μm) resin comes from Eichrom, Lisle IL. The cyclic chelating ligand1,4,7,10-tetraazacyclodo-decane-1,4,7,10-tetraacetic acid (DOTA) was purchased from Macrocyclics, Dallas TX. The acyclic chelating ligand diethylenetriamine pentaacetic acid (DTPA) was purchased from Acros Organics, Geel, Belgium. Sodium acetate (NaOAc) was purchased from Fisher Scientific, Pittsburg PA. Both chelating ligands and NaOAc were dissolved in 18 MΩ-cm deionized (DI) water and mixed with Chelex 100 from Sigma-Aldrich for trace metal purification. Scandium foil (99.9%) was purchased from Alfa Aesar, Ward Hill MA. A 50 ppm multi-element standard for calibration and Agilent’s 4200 Microwave Plasma Atomic Emission Spectroscopy (MP-AES) system come from Agilent Technologies, Santa Clara CA.
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2

Synthesis of Cy5.5-DOTA Conjugate

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All chemicals and solvents were obtained from commercial sources and used without further purification. Sulfo-N-hydroxysulfosuccinimide (Sulfo-NHS) and ethyl-(dimethylaminopropyl) carbodiimide (EDC) were purchased from Sigma-Aldrich. 1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) was obtained from Macrocyclics, Inc. Cy5.5-Mal was purchased from GE healthcare.
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3

Lipid-based Nanoparticle Formulation Protocol

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POPC; DSPC; DOTAP; DSTAP; cholesterol; and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(PEG)-2000] (DSPE-PEG2000) were purchased from Avanti Polar Lipids Inc. (Alabaster, AL, USA). 3H-cholesterylhexadecyl ether (3H-CHE) was from PerkinElmer (Skovlunde, Denmark). 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) was purchased from Promega. DQ gelatin was from Life Technologies (Taastrup, Denmark). 9-Fluorenylmethoxycarbonyl (Fmoc) amino acids and O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU) were purchased from GL Biochem (Shanghai, China) or Bachem AG (Bubendorf, Switzerland). TentaGel PAP2000 resin was purchased from Rapp Polymere GmbH (Tuebingen, Germany). DiI was purchased from Thermo Fisher Scientific. Oxaliplatin (L-OHP) was from Lianyungang Guiyuan Chempharm Co. Ltd. (Jiangsu, China). 1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) was purchased from Macrocyclics (Dallas, TX, USA). All other chemicals were from Sigma-Aldrich (Brøndby, Denmark) and of analytical grade.
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