Ferrozine 

Ferrozine is a spectrophotometric reagent for iron ions, can react with divalent Fe to form a stable magenta complex species. The complex has an absorption peak at 562 nm^[1][2]. Ferrozine-based colorimetric assays can quantify iron in cells^[3]

Guidelines (The following is our recommended protocol. This protocol is provided for reference only and should be modified according to your specific needs).
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.

492.46

C₂₀H₁₃N₄NaO₆S₂

562

69898-45-9

Solid

Light yellow to yellow

O=S(C1=CC=C(C2=NC(C3=NC=CC=C3)=NN=C2C4=CC=C(S(=O)(O[Na])=O)C=C4)C=C1)(O)=O

Room temperature in continental US; may vary elsewhere.

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

• [1]. Ferrozine-A New Spectrophotometric Reagent for Iron. ANALYTICAL CHEMISTRY, VOL. 42, NO. 7, JUNE 1970. 779-781

  [2]. E Viollier, et al. The ferrozine method revisited: Fe(II)/Fe(III) determination in natural waters.

  [3]. Riemer  et al.Colorimetric ferrozine-based assay for the quantitation of iron in cultured cells. Anal Biochem. 2004 Aug 15;331(2):370-5.  [Content Brief]