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3 protocols using tetramethylthiuram disulfide

1

Disulfiram and Copper Compounds in NSCLC

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The non-small cell lung cancer (NSCLC) A549 (CCL-185) and H23 (CRL-5800) cell lines were purchased from ATCC (Middlesex, UK). Copper chloride (CuCl2, Cu), disulfiram (DS), diethyldithiocarbamate (DDC), S-methyl-N,N-diethyldithiocarbamate (S-Me-DDC), tetramethylthiuram disulfide (TMDS), 2-hydroxy-dithiobenzoic acid (HDTA), 4-imidazoledithiocarboxylic acid (IDTA), 2,4,6-Trimercaptotriazine (TMT) and poly-2-hydroxyethyl methacrylate (poly-HEMA) were purchased from Sigma (Dorset, UK). Bis(N,N-diethyldithiocarbamate)-copper(II) (Cu-DDC) was from Santa Cruz (Dallas, TX, USA). Annexin V kit was from Roche Applied Sciences (Burgess Hill, UK). Fc OxyBURST Assay Reagents was purchased from Invitrogen, Molecular Probes (Waltham, MA, USA). ALDEFLUOR kit was from StemCell Tech (Durham, NC, USA). FITC mouse Anti-Human CD44 from BD Biosciences (Oxford, UK).
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2

Synthesis and Characterization of Gold Nanoparticles

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Tetrachloroauric acid (30% in HCl, 99.99%), 2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol, sodium borohydride (≥98%), ascorbic acid (≥99%), silver nitrate (≥99%), 7-mercapto-4-methylcoumarin (≥97%), (3-aminopropyl)triethoxysilane (APTES) (≥98%), norfloxacin (≥98%), tetramethylthiuram disulfide (≥97%), ethanol (≥99.8%), toluene (≥99.7%), hydrochloric acid (≥37%), nitric acid (≥65%), sulphuric acid (≥95%), hydrogen peroxide (30 wt%) and ammonium hydroxide solution (≈29%) were purchased from Sigma Aldrich.
Microscopy cover glass slides 21 mm × 26 mm have been obtained from DEL Chimica and PDMS (Sylgard 184) was purchased from Dow Chemical.
Water was deionized and double distilled (ddH2O).
All the glassware that came in contact with nanoparticles was always cleaned with aqua regia for 15 min, then washed three times with bi-distilled water for 3 min under sonication and dried in an oven for 1 h at 140 °C.
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3

Allergy-Inducing Chemicals Protocol

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4-Ethoxymethylene-2-phenyl-2-oxazolin-5-one (oxazolone), 2,4dinitrochlorobenzene, acetaminophen, 3-aminophenol, cinnamic aldehyde, isoeugenol, citral, tetramethylthiuram disulfide, 2-methoxy-4-methylphenol, resorcinol, eugenol, geraniol, cinnamic alcohol, benzalkonium chloride, vanillin, lactic acid, N,N-diethyl-3-methylbenzamide and 4-aminoacetanilide were obtained from Sigma Aldrich (Taufkirchen, Germany), whereas N,N'-bis(4-aminophenyl)-2,5-diamino-1,4-quinone-diimine (Bandrowski's base) was purchased from ICN Biomedicals (Aurora, OH, USA). Sodium dodecyl sulfate and dimethyl sulfoxide (DMSO) were purchased from Carl Roth (Karlsruhe, Germany). 4-Acetamidoacetanilide was synthesized as described (Kawakubo et al., 2000) . 4-Amino-2-methyl-acetanilide, 4-amino-3-methyl-acetanilide and 2,5-diacetaminotoluene were kind gifts from C. Goebel (P&G, Darmstadt, Germany).
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