BioReagent BioReagent for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C,Protected from light,Room temperature Ships Wet ice Check lot-specific COA for exact specifications.
SDS, COA, datasheet, and spec sheet available for download. Lot-specific COA accessible via lot number lookup.
Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
Iron is an essential trace element in the human body, with a total content of approximately 3270 mg. It is widely distributed: 67.6% of iron is found in hemoglobin as a prosthetic group, involved in oxygen transport; 2.59% and 4.15% are present in bone and myoglobin, respectively; and storage iron accounts for about 25.37%. Serum iron is bound to transferrin in the form of ferric ions (Fe³⁺). Therefore, measuring serum iron requires first dissociating Fe³⁺ from transferrin.
Assay Principle
This assay utilizes a spectrophotometric method with Ferrozine as the chromogen. In an acidic medium, serum iron bound to transferrin is released. Iron in other samples is also liberated under acidic conditions. The released iron is then reduced to ferrous ions (Fe²⁺) by a reducing agent (present in the assay buffer). The Fe²⁺ subsequently reacts with Ferrozine to form a magenta-colored complex. The absorbance at 562 nm is measured using a spectrophotometer. This method is suitable for detecting iron content in samples such as serum, plasma, and tissues. As this direct detection method is used, a serum blank should be included to correct for the sample's inherent color. The iron content is calculated according to the formula. This kit shows a good linear relationship up to 140 μmol/L. Triglycerides ≤ 3.39 mmol/L and bilirubin ≤ 171 μmol/L basically do not interfere with this method. This kit is for research use only and not suitable for clinical diagnosis or other purposes.
| I1505783 | Component | 50T | Storage |
| I1505783A | Iron Standard (100 μg/mL) | 1 mL | 2-8℃. Store in the dark. |
| I1505783B | Iron Standard Diluent | 10 mL | RT |
| I1505783C | Fe Assay Buffer | 60 mL | 2-8℃ |
| I1505783D | Ferrozine Chromogenic Solution | 3 mL | 2-8℃. Store in the dark. |
| I1505783E | ddH₂O | 10 mL | RT |
Required Materials Not Provided
Centrifuge tubes or test tubes, Spectrophotometer, Cuvettes
Experimental Procedure
1. (Optional) Sample Preparation
1.1 Plasma/Serum Samples
Prepare plasma or serum by conventional methods.
Store at -20°C for Fe detection.
1.2 Cell or Tissue Samples
Homogenize an appropriate amount of cells or tissue.
Centrifuge at low speed and collect the supernatant.
Store at -20°C for Fe detection.
1.3 High-Concentration Samples
If the sample contains a high concentration of Fe, dilute it with ddH₂O. Do not use regular distilled water for dilution.
Note: (Optional) After preparation, determine the protein concentration using a BCA Protein Assay Kit to calculate Fe content per mg protein.
2. Preparation of Iron Standard Working Solution
Dilute the Iron Standard (100 μg/mL) with Iron Standard Diluent at a 1:49 ratio to prepare the Iron Standard Working Solution (2 μg/mL).
Store at 4°C protected from light; stable for 3 months.
3. Fe Assay Setup
Use dry test tubes or disposable sterile polyethylene centrifuge tubes cleaned with dilute hydrochloric acid and rinsed with deionized water.
Set up Blank, Standard, and Test tubes according to the table below.
Add reagents in the specified order, avoiding bubbles.
If the sample's iron concentration is too high, reduce the sample volume or dilute appropriately before assaying. Duplicate tubes are recommended.
| Reagent (μL) | Blank Tube | Standard Tube | Test Tube |
| ddH₂O | 0.45 | — | — |
| Iron Standard Working Solution (2 μg/mL) | — | 0.45 | — |
| Sample | — | — | 0.45 |
| Fe Assay Buffer | 1.2 | 1.2 | 1.2 |
Mix well. Read absorbance at 562nm, zeroed with the Blank tube. Record for Test tube (This is the Serum Blank,
Ferrozine Chromogenic Solution | 0.05 | 0.05 | 0.05 |
4. Fe Measurement
Mix well.
Incubate at room temperature for 15 minutes or at 37°C for 10 minutes.
Zero the instrument with the Blank tube.
Unit Conversions:
Iron Standard (2 μg/mL) = Iron Standard (2 mg/L) = Iron Standard (35.8 μmol/L)
μg/dL = μmol/L / 0.179
μmol/L = 0.179 × μg/dL = 17.9 × μg/mL
Reference Interval:
Healthy Adults Serum Iron:
Male: 11 ~ 30 μmol/L (60 ~ 170 μg/dL)
Female: 9 ~ 27 μmol/L (50 ~ 150 μg/dL)
6. Standard Curve Reference
*(Placeholder for standard curve graph and data, typically showing absorbance vs. concentration for standards like 0, 0.5, 1, 2, 4, 6, 8, 10 μg/mL, measured at 562 nm)*
*Note: Standard curves may vary due to differences in instruments and operational techniques. Empirical data suggests deviations in the standard curve at iron standard concentrations below 0.1 μg/mL and above 40 μg/mL.*
Precautions
Hemolyzed samples interfere with the assay; avoid using them if possible.
If the sample concentration is too high, dilute with distilled water and re-assay, multiplying the result by the dilution factor.
Use high-purity deionized water during the experiment; do not use ordinary distilled water.
Glassware should be soaked in 10% hydrochloric acid for 24 hours and rinsed with deionized water before use.
Avoid contact with iron objects to prevent iron contamination.
The standard color is stable for 24 hours. The serum color is stable for 30 minutes; the color intensifies slowly over time, so measurement must be completed within 1 hour.
The value 0.97 is a volume correction factor.
The within-run coefficient of variation (CV) for this method is ≤ 3.1%; the between-run CV is ≤ 2.6%.
Use reagents promptly after opening to avoid affecting subsequent experimental results.
For your safety and health, wear lab coats and disposable gloves during operation.
Comprehensive hazard, handling, storage, and regulatory compliance document.
Download SDS →Lot-specific quality data. Enter your lot number to retrieve the exact COA.
Look up COA →Full quality attributes and acceptance criteria for this grade.
View spec sheet →Find and download the COA for your product by matching the lot number on the packaging.
| Lot Number | Certificate Type | Fecha | Articulo |
|---|---|---|---|
 ZJ26F0636792 | Certificate of Analysis | Jun 26, 2026 | I1505783 |
| C2624378 | Certificate of Analysis | Mar 24, 2026 | I1505783 |
| L2511008 | Certificate of Analysis | Dec 11, 2025 | I1505783 |
| Sensibilidad | Light-sensitive |
|---|
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