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4 protocols using copper 2 chloride dihydrate

1

Multimodal Cellular Imaging Protocol

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Copper(II) chloride dihydrate
(CuCl2·2H2O), gadolinium(III) chloride
hexahydrate (GdCl3·6H2O), sodium hydroxide
(NaOH), bovine serum albumin, sodium sulfide pentahydrate (Na2S·5H2O), methylene blue, and Hoechst 33342
solution were purchased from Fujifilm Wako Pure Chemical Co. (Osaka,
Japan). Sulfo-NHS-Cy5.5 ester was obtained from Abcam (Cambridge,
UK). Cyclo(RGDfk) and 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA)
were obtained from Cayman Chemical (Michigan, USA). Sulfo-NHS-acetate
was purchased from BroadPharm, California (USA). 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
hydrochloride (EDC), N-hydroxysulfosuccinimide (sulfo-NHS),
Alexa Fluor 488 phalloidin, LysoTracker Green DND-26, calcein AM,
and propidium iodide (PI) were obtained from Thermo Fisher Scientific
(Massachusetts, USA). All other chemicals of analytical reagent grade
were obtained from qualified reagent suppliers.
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2

Transition Metal Chloride Synthesis and Analysis

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Nickel chloride (NiCl2), zinc chloride (ZnCl2), cobalt chloride (CoCl2), copper (II) chloride dihydrate (CuCl2·2H2O), iron (II) chloride tetrahydrate (FeCl2·4H2O), magnesium chloride hexahydrate (MgCl2·6H2O), manganese (II) chloride tetrahydrate (MnCl2·4H2O), lipopolysaccharides (LPS) from Escherichia coli O111, and 30% (w/v) H2O2 were purchased from Wako Pure Chemical Industries (Osaka, Japan). Chlorazol Black and TAK-242 were purchased from Sigma-Aldrich Co. (St. Louis, MO) and Calbiochem-Merck Millipore (Darmstadt, Germany), respectively. Newport GreenTM DCF diacetate was purchased from Invitrogen (Carlsbad, CA) and the Ni wire (purity 99.98%, diameter 0.8 mm) from Nilako (Tokyo, Japan). HNO3 (69% (w/w)) was purchased from Kanto Chemical Co., Inc. (Tokyo, Japan).
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3

Synthesis of Metal-Chelated Antioxidants

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Lauric
acid (CH3(CH2)10COOH), magnesium(II)
chloride hexahydrate, manganese(II)
chloride tetrahydrate, cobalt(II) chloride hexahydrate, nickel(II)
chloride hexahydrate, copper(II) chloride dihydrate, zinc chloride,
and phosphate buffer (pH 6) were purchased from Wako Pure Chemical
Industries Ltd. (Osaka, Japan). Calcium(II) chloride dihydrate was
obtained from Sigma-Aldrich Co. LLC (St. Louis). Sodium hydroxide
was purchased from Kanto Chemical Co. Inc. (Kanagawa, Japan). DMPO
was purchased from Labotech Inc. (Tokyo, Japan).
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4

Purification and Preparation of Reagents for Polymer Synthesis

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4-Chloromethylstyrene (CMS, >90%, TCI) was purified by column chromatography with basic aluminum oxide (Aldrich) to remove the stabilizer before use. Divinylbenzene (DVB, m- and p-mixture, >50%, TCI) was extracted from a sodium hydroxide aqueous solution (1 M) to remove the polymerization inhibitor before use. 2,2′-Azobis(isobutyronitrile) (AIBN, >98%, TCI) was crystallized from ethanol. Methoxy polyethylene glycol acrylate AM-90G (MGA, –(OCH2–CH2)9) was provided by Shin-Nakamura Chemical Co., Ltd. Potassium carbonate (K2CO3, >99.0%, TCI), dimethylacetamide (DMAc, Super dehydrated, >99.5%, Wako), polyvinylpyrrolidone K25 (PVP, Wako), hydrochloric acid (5 M, Wako), ethanol (>99.5%, Wako), theophylline (Wako), palladium chloride (>99.0%, Wako), hydrogen hexachloroplatinate(iv) hexahydrate (H2PtCl6·6H2O, >98.5%, Wako), copper(ii) chloride dihydrate (CuCl2·2H2O, >99.0%, Wako), bromobenzene (>99.0%, TCI), phenylboronic acid (TCI), chloroform-D1 (CDCl3 >99.8%, 0.03% TMS) and other reagents were used as received without further purification.
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