Fe2+ chloride (Sigma-Aldrich, 372,870) was dissolved in water to a final concentration of 10 μM. The tested compounds were added to 1 ml of Fe2+ chloride solution with a final concentration of 40 μM and incubated at room temperature for 10 min. Next, FerroZine Iron Reagent (Sigma-Aldrich, 160,601) was added with a final concentration of 20 μM and incubated at room temperature for 1 h [67 (link)]. The absorbance at 562 nm was measured using a microplate reader (Thermo Scientific, Varioskan Flash). Results were normalized to DMSO control group (DMSO vehicle; set as 100%).
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Ferrous Iron Reduction Assay
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Corresponding Organization : The University of Texas Southwestern Medical Center
Other organizations : University of Michigan–Ann Arbor, Centre de Recherche des Cordeliers, Université Paris Cité, Inserm, Institut Universitaire de France, Sorbonne Université, Hôpital Européen, Hôpital Européen Georges-Pompidou, Assistance Publique – Hôpitaux de Paris, Institut Gustave Roussy
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Variable analysis
independent variables
- Tested compounds
- Absorbance at 562 nm
- Fe2+ chloride concentration (10 μM)
- Fe2+ chloride solution volume (1 ml)
- Tested compounds concentration (40 μM)
- Incubation time (10 min)
- FerroZine Iron Reagent concentration (20 μM)
- Incubation time with FerroZine Iron Reagent (1 h)
- DMSO control group
- Not explicitly mentioned
- DMSO control group
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