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#DIFE-250 QuantiChrom™ Iron Assay Kit
Product name : QuantiChrom™ Iron Assay Kit
Catalog number : DIFE-250
Supplier name : Bioassays
Data sheet: Ask more or other datasheet now !
About this Product :QuantiChrom™ Iron Assay Kit antibody storage GENTAUR recommends for long therm storage to freeze at -24 C. For short time storage up to 30 days we suggest fridge storage at 1 to 10 C. Prevent multiple freeze taw cycles of QuantiChrom™ Iron Assay Kit.
More Details about
Iron level in blood is a reliable diagnostic indicator of various disease
states. Increased levels of iron concentration in blood are associated with
blood loss, increased destruction of red blood cells (e.g. hemorrhage) or
decreased blood cell survival, acute hepatitis, certain sideroachrestic
anemias, ingestion of iron-rich diets, defects in iron storage (e.g.
pernicious anemia). Decreased levels of blood iron may result from
insufficient iron ingestion from diets, chronic blood loss pathologies, or
increased demand on iron storage as during normal pregnancy.
Simple, direct and automation-ready procedures for measuring iron
concentrations find wide applications in research, drug discovery and
environmental monitoring. BioAssay Systems' iron assay kit is designed
to measure total iron directly in serum without any pretreatment. The
improved method utilizes a chromogen that forms a blue colored complex
specifically with Fe2+. Fe3+ in the sample is reduced to Fe2+, thus allowing
the assay for total iron concentration. The intensity of the color, 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. The procedure involves addition of a
single working reagent and incubation for 40 min. Can be readily
automated as a high-throughput assay for thousands of samples per day.
Improved reagent stability and versatility. The optimized formulation has
greatly enhanced reagent and signal stability. Cuvette or 96-well plate assay.
Low interference in biological samples. No pretreatments are needed.
Assays can be directly performed on serum samples.
Direct Assays: iron in biological samples (e.g. serum).
Drug Discovery/Pharmacology: effects of drugs on iron metabolism.
Environmental Monitoring: iron in soil extracts, mineralized samples.
KIT CONTENTS (250 tests in 96-well plates)
Reagent A: 50 mL Reagent B: 4 mL Reagent C: 4 mL
Iron Standard: 1 mL 10 mg/dL Fe2+
Storage conditions. 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.
Precautions: reagents are for research use only. Normal precautions for
laboratory reagents should be exercised while using the reagents. Please
refer to Material Safety Data Sheet for detailed information.
Note: (1). Iron chelators (e.g. EDTA) interfere with this assay and should
be avoided in sample preparation. (2). Serum or plasma samples should
be clear and free of precipitates or turbidity. If not, centrifuge or filter to
clarify samples prior to assay. (3).This kit can be applied to measure Fe2+
(vs. total iron) content. Prepare Working Reagent by mixing 20 vol of
Reagent A, 1 vol of water and 1 vol Reagent C (no reductant in the
Working Reagent). The procedure is the same as described for the total
Procedure using 96-well plate:
1. Standards. Prepare 400 μL 1000 μg/dL Premix by mixing 40 μL 10
mg/dL standard and 360 μL distilled water. Dilute standards as follows.
No Premix + H2O Vol (μL) Fe (μg/dL)
1 100μL + 0μL 100 1000
2 80μL + 20μL 100 800
3 60μL + 40μL 100 600
4 40μL + 60μL 100 400
5 30μL + 70μL 100 300
6 20μL + 80μL 100 200
7 10μL + 90μL 100 100
8 0μL + 100μL 100 0
Transfer 50 μL diluted standards and 50 μL sample into a clear flat
bottom 96-well plate. For serum/plasma samples, it is recommended
to run a sample blank (i.e. a 50 μL sample in a separate well).
2. Prepare enough Working Reagent by mixing 20 volumes of Reagent A, 1
volume Reagent B and 1 volume Reagent C. Fresh reconstitution is
recommended. Equilibrate to room temperature before assay.
Add 200 μL Working Reagent to Standards and Samples wells. (For
serum/plasma samples which require a Sample Blank Control, add
200 μL Reagent A to the Sample Blank wells). Tap plate to mix.
3. Incubate 40 min at room temperature and read optical density at 510-
630nm (peak absorbance at 590nm).
Procedure using cuvette:
1. Prepare standards as in 96-well assay. Set up centrifuge tubes
labeled Standards and Samples. Transfer 250 μL standards and
samples to tubes.
2. Add 1000 μL Working Reagent to all tubes. Mix by vortexing. Incubate
40 min at room temperature.
3. Transfer to cuvettes and read OD at 590nm (510nm-630nm).
Subtract OD of “0 μg/dL Fe” from all other standard OD values and plot
the OD against standard iron concentrations. Determine the slope using
linear regression fitting. Iron concentration of the sample is calculated as
Where ODBLANK is OD values of the water blank (Standard #8), or
Sample Blank, if a sample blank is used (e.g. serum or plasma). Typical
serum iron values: 70-180 μg/dL.
Conversions: 1 mg/dL Fe equals 179 μM, 0.001% or 10 ppm.
MATERIALS REQUIRED, BUT NOT PROVIDED
Pipeting devices and accessories.
Procedure using 96-well plate:
Clear bottom 96-well plates (e.g. Corning Costar) and plate reader.
Procedure using cuvette:
Cuvets and spectrophotometer for measuring OD at 510-630nm.
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WP1566: Citrate cycle (TCA cycle)
WP1596: Iron Homeostasis
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WP1680: Oxidative phosphorylation
WP2007: iron metabolism in placenta
WP407: Kit Receptor Signaling Pathway
WP548: neural crest development
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