Description
Iron Assay Kit (BA0040) (BA0040)
The Iron Assay Kit (SKU: BA0040) provides a simple, direct and automation-ready method for the quantitative colorimetric determination of total iron in biological and environmental samples. Iron levels in blood serve as a reliable diagnostic indicator of numerous disease states, with raised levels associated with haemolysis, acute hepatitis and iron-rich diets, and reduced levels linked to insufficient dietary intake or chronic blood loss. The assay measures total iron directly in serum without any pretreatment, using a chromogen that forms a blue-coloured complex specifically with Fe2+. Ferric iron present in the sample is reduced to Fe2+, allowing the determination of total iron concentration. The intensity of the resulting colour, measured at 590nm, is directly proportional to the iron concentration in the sample. This makes the kit well suited to research, drug discovery and environmental monitoring applications.
| Product Name: | Iron Assay Kit (BA0040) |
| SKU: | BA0040 |
| Detection Method: | Colorimetric |
| Detection Range: | 27 µg/dL (4.8 µM) to 1,000 µg/dL (179 µM) iron in 96-well plate assay |
| Sample Type: | Serum, plasma, soil extracts and mineralised samples |
| Species Reactivity: | All |
| Assay Time: | 40 minutes |
| Kit Size: | 250 Assays |
| Equipment Required: | Microplate reader |
| Storage: | The kit is shipped at room temperature. Store Reagent A at room temperature and other reagents at 4°C. |
| Shelf Life: | 12 months after receipt |
| Shipping: | Room Temperature |
This kit measures total iron directly in serum without pretreatment. The improved method utilises a chromogen that forms a blue-coloured complex specifically with Fe2+, with Fe3+ in the sample reduced to Fe2+ to allow the assay of total iron. The intensity of the colour, measured at 590nm, is directly proportional to the iron concentration in the sample.
- Sensitive and accurate: linear detection range 27 µg/dL (4.8 µM) to 1,000 µg/dL (179 µM) iron in 96-well plate assay
- Simple and high-throughput: addition of a single working reagent and incubation for 40 minutes, readily automated for thousands of samples per day
- Improved reagent stability and versatility in cuvette or 96-well plate format
- Low interference in biological samples: no pretreatments required, assays performed directly on serum
- Direct assays: iron in biological samples (e.g. serum)
- Drug discovery/pharmacology: effects of drugs on iron metabolism
- Environmental monitoring: iron in soil extracts and mineralised samples
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Standards: prepare 400 µL 1000 µg/dL Premix by mixing 40 µL 10 mg/dL standard and 360 µL distilled water, then dilute as shown in the table. Transfer 50 µL diluted standards and 50 µL sample into a clear flat-bottom 96-well plate. For serum/plasma, a sample blank (50 µL sample in a separate well) is recommended. |
| 2 | Prepare Working Reagent by mixing 20 volumes Reagent A, 1 volume Reagent B and 1 volume Reagent C (fresh reconstitution recommended; equilibrate to room temperature). Add 200 µL Working Reagent to standards and samples wells (add 200 µL Reagent A to sample blank wells). Tap plate to mix. |
| 3 | Incubate 40 minutes at room temperature and read optical density at 510-630nm (peak absorbance at 590nm). |
Subtract the OD of the 0 µg/dL Fe standard from all other standard OD values and plot OD against standard iron concentrations. Determine the slope by linear regression. Iron concentration = (ODSAMPLE − ODBLANK) / Slope (µg/dL), where ODBLANK is the water blank (Standard #8) or sample blank if used. Typical serum iron values: 70-180 µg/dL. Conversion: 1 mg/dL Fe equals 179 µM, 0.001% or 10 ppm.
| Component | Quantity | Storage |
| Reagent A | 50 mL | Room temperature |
| Reagent B | 4 mL | 4°C |
| Reagent C | 4 mL | 4°C |
| Iron Standard (10 mg/dL Fe2+) | 1 mL | 4°C |
