Ferrozine (solution)

Colorimetric ferrozine-based assay for the quantitation of iron in cultured cells^[3]
1: Preparation of Reagents
?1.1: Iron Releasing Reagent: Prepare a mixture of 1.4 M HCl and 4.5% KMnO4.
?1.2: Iron Detection Reagent: Prepare a solution containing 6.5 mM Ferrozine, 6.5 mM neocuproine(HY-W004563), 2.5 M ammonium acetate, and 1 M ascorbic acid.
2: Cell Treatment: Treat cultured cells with different concentrations of Ferric Ammonium Citrate (FAC) (HY-B1645) and incubate for 24 hours. After treatment, wash the cells with PBS and collect them.
3: Cell Lysis and Iron Release
?3.1: Cell Lysis: Lyse cells using 50 mM NaOH, typically agitating at room temperature for 2 hours.
?3.2: Iron Release: Add the iron releasing reagent to the lysate and then incubate at 60°C for 2 hours.
4: Colorimetric Determination of Iron Content
?4.1: Iron Detection Reaction: Mix the lysate with the iron detection reagent and react for 30 minutes.
?4.2: Absorbance Measurement: Measure the absorbance at 550 nm using a microplate reader.
5: Data Analysis
?5.1: Standard Curve: Prepare standard solutions with known concentrations of iron, measure their absorbance at 550 nm, and establish a standard curve.
?5.2: Iron Content Calculation: Calculate the iron content of the samples by comparing their absorbance to the standard curve.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.