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

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Ferrozine Chemical Structure

Ferrozine Chemical Structure

CAS No. : 69898-45-9

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Description

Ferrozine is a spectrophotometric reagent for iron, 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]

In Vitro

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.

Molecular Weight

492.46

Formula

C20H13N4NaO6S2

CAS No.
Appearance

Solid

Color

Light yellow to yellow

Excitation (Ex)

562

SMILES

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

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

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)

Solvent & Solubility
In Vitro: 

DMSO : 43.75 mg/mL (88.84 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.0306 mL 10.1531 mL 20.3062 mL
5 mM 0.4061 mL 2.0306 mL 4.0612 mL
View the Complete Stock Solution Preparation Table

* 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.

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Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

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In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

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g

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(per animal)

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Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration: mg/mL
Purity & Documentation

Purity: 99.96%

References

Complete Stock Solution Preparation Table

* 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.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 2.0306 mL 10.1531 mL 20.3062 mL 50.7655 mL
5 mM 0.4061 mL 2.0306 mL 4.0612 mL 10.1531 mL
10 mM 0.2031 mL 1.0153 mL 2.0306 mL 5.0766 mL
15 mM 0.1354 mL 0.6769 mL 1.3537 mL 3.3844 mL
20 mM 0.1015 mL 0.5077 mL 1.0153 mL 2.5383 mL
25 mM 0.0812 mL 0.4061 mL 0.8122 mL 2.0306 mL
30 mM 0.0677 mL 0.3384 mL 0.6769 mL 1.6922 mL
40 mM 0.0508 mL 0.2538 mL 0.5077 mL 1.2691 mL
50 mM 0.0406 mL 0.2031 mL 0.4061 mL 1.0153 mL
60 mM 0.0338 mL 0.1692 mL 0.3384 mL 0.8461 mL
80 mM 0.0254 mL 0.1269 mL 0.2538 mL 0.6346 mL
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Ferrozine
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