Compounds were plated at 10 μM in DMF in 96-well plates. Reactions were initiated by adding citrate buffer (20 mM sodium citrate pH 6, 50 mM sodium chloride), 0.1 U/mL horseradish peroxidase solubilized in citrate buffer (P8250, Millipore Sigma), and 50 μM Amplex Red solubilized in DMSO (A12222, ThermoFisher Scientific) in a 100 μL volume reaction. Control wells received 10 U/mL catalase (C9322, Millipore Sigma). Fluorescence readings were taken with a 528 nm excitation/590 nm emission filter set using a Biotek Synergy 4 plate reader.
Synergy 4 plate
Manufactured by Agilent Technologies
The Synergy 4 plate is a multi-mode microplate reader designed for a variety of applications in life science research laboratories. It features a modular design, allowing the integration of different detection modes such as absorbance, fluorescence, and luminescence measurements. The Synergy 4 plate can be used to perform various assays, including cell-based, biochemical, and molecular biology experiments.
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Lab products found in correlation
3 protocols using synergy 4 plate
1
Horseradish Peroxidase Activity Assay
2
Measuring Intracellular ROS in HepG2 Cells
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H2DCFDA was used to measure ROS production. Upon exposure to ROS, the chemical is converted to the highly fluorescent DCF molecule. After washing with HBSS, HepG2 cells cultured in SeedEZ scaffolds in PerfusionPal or in a multi-well plate were incubated with 5 mM of H2DCFDA (Invitrogen, Carlsbad, CA, United States) at room temperature for 20 min. The scaffolds were transferred to a black-wall 24-well plate and the fluorescence (485 nm excitation/530 nm emission) was read on a BioTek Synergy 4 plate reader.
3
Thioflavin T Fluorescence Assay
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Assays were carried out as described by LeVine [30 (link)]. Thioflavin T (Sigma Chemical, St. Louis, MO) was dissolved in 50mM glycine-NaOH, pH 8.5 and diluted to 10 μM when mixed with proteins. The fluorescence was read in a Shimadzu RF-5301PC spectrofluorophotometer in spectrum mode, with the excitation filter at 450+/-5 nm and the emission filter at 485+/-5 nm or using a Biotek Synergy 4 plate reader (384 well; 50 μl per well; triplicate wells) with excitation at 450 nm and emission at 490 nm.
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