1420 multilabel counter victor 3v plate reader

Binding assays were conducted as described (59 (link)). In short, all of the Fluorescein Amidite–labeled G-rich RNA substrates (Table S1) were suspended in a G4-folding buffer containing 100 mM KCl, heated at 95 °C for 6 min and slowly cooled to room temperature. The folded G4 substrates (2 nM final concentration, unless stated otherwise) were preincubated in binding buffer (25 mM MOPS, pH 6.5; 100 mM KCl, 0.1 mM EDTA, 2 mM β-mercaptoethanol, 0.1 mg/ml bovine serum albumin in DEPC-treated water) with increasing concentrations of NS3 in a 96-well plate for 30 min at 37 °C. A PerkinElmer 1420 multilabel counter victor ³V plate reader was used to determine the polarization of NS3-G4 complex with 485 nm and 535 nm excitation and emission wavelengths, respectively. The polarization measurements were converted to fluorescence anisotropy values, which were then fit to a quadratic equation (unless otherwise stated) to acquire dissociation constants (K_d).

Many common cytotoxicity assays [MTT (3-[4,5-dimethyl-2-thiazol]-2,5-diphenyl-2H-tetrazolium bromide), MTS (4-[5-[3-(carboxymethoxy)phenyl]-3-(4,5-dimethyl-1,3-thiazol-2-yl)tetrazol-3-ium-2-yl]benzenesulfonate), alamar blue (resazurin), neutral red (3-amino-7-dimethylamino-2 methylphenazine hydrochloride), ATP and simple visual examination of the cells] have been used by our laboratory seeking to avoid or minimize interferences from the nanomaterials themselves. After 72 h of culture with various nanomaterials, cytotoxicity assays based on MTT (Sigma-Aldrich, St Louis, MO), MTS (Promega, Madison, WI) and alamar blue (Cell Tier-Blue, Promega, Madison, WI) were performed in accordance with the enclosed kit directions. Alamar blue and MTS were used for all nanomaterial cytotoxicity experiments except for CeO₂ Q (MTT only was used). A PerkinElmer 1420 Multilabel Counter Victor³V plate reader was used for all cytotoxicity assays. Cytotoxicity assays results were always checked with each other and versus visual assessment of the cells to ensure the cytotoxicity assays were functioning properly.