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Iron Assay kit LS – Ferrozine method | Metallo Assay LS

Date:June 11 2020Web Page No:80480

Iron Assay kit LS - Ferrozine method

Metallo Assay Iron Assay Kit LS (Ferrozine Method) can measure Iron ion (Fe2+/Fe3+ ) in the biological samples such as Plasma, Serum, Urine, and wide range of samples by microplate reader.

Iron is contained in various enzymes as an important constituent element. Iron in the blood are all combined to transferrin, and transported to erythroblast and tissues for synthesis of globin proteins such as myoglobin and hemoglobin which requests iron. Iron is essential for generation of oxigen transporting proteins. Deficient of such proteins cause hypoferric anemia, chronic bleeding anemia, and infectious anemia. Increase of transferrin and high concentration of iron are reported in hepatitis and liver cirrhosis. Increase of iron is reported in Hypoplastic anemia and malignant anemia.

Features

  • Using microplate reader for assay, no need to use Atomic Absorption Spectrometry or ICP-OES Optical Emission Spectrometry, which have been mainly used so far.
  • Good correlation with the result of Atomic Absorption Spectrometry and ICP-OES.
  • High throughput measurement by using microplate reader.
  • All animal species sample can be measured.
  • Assay time : just 10 minutes!
  • 1 point calibration – do not need multiple point measurement to make standard curve (r2=0.9999).
  • Kit does not contain any hazardous components like cyanide or azide as preservative.
  • Samples : Serum, Plasma, Urine, Saliva, Cell lysate, Tissue extract, Hair extract, Water, etc.
  • In combination with UIBC Assay kit LS Bathophenanthroline method (#UIB02AE), TIBC (Total Iron Binding Capacity) can be measured.

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Assay Principle

This kit measures iron concentration by color change caused by chelate complex formulation of Ferrozine and Iron. Iron combined to transportation protein such as transferrin is denatured by weak acid and denaturant then formulates Iron-Ferrozine chelate complex. Measuring this complex by 560 nm wavelength and obtain iron concentration.

Assay Principle of Ferrozine method Iron Assay kit LS

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Sample Preparation

Sample preparation of Ferrozine method Iron Assay kit LS
  • Samples which do not require preparation : e.g. Serum, Plasma and Urine
    * EDTA treated sample is not suitable for this assay.
    * Pooled urine should add HCl for storing. When assay, adujust sample pH 2 to 3.

  • Samples which require acid extraction : e.g. Cell Lysate, Tissue Homogenate and Other liquid samples
    * Measurable chemical species is limited as follows:
      ・Free iron
      ・Iron binds to protein (Protein-Fe2+, e.g. albumin, metallothionein)
    * Lysis buffer contains EDTA is not suitable for this assay.
    * Organoiron and Heme iron requires acidolysis.

  • Microwave method and samples require acidolysis : e.g. Samples contain strong chelate agents such as EDTA, Organoiron (in the case of single bond -C-Fe-C-), Iron in cyclic ligand (Fe-Porphyrin)
    * After acidolysis, adjust sample pH in 2 to 3.

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Handling Procedure

(1)Apply DW, Standard and Sample (40 uL) to 96 well plate. Handling procedure 1 of Ferrozine method Iron Assay kit LS
(2)Add R-A buffer (200 uL). Handling procedure 2 of Ferrozine method Iron Assay kit LS
(3)Incubate at room temperature for 5 minutes. Handling procedure 3 of Ferrozine method Iron Assay kit LS
(4)Read the absorbance at 560 nm (main) as OD1. Handling procedure 4 of Ferrozine method Iron Assay kit LS
(5)Add R-R Chelate Color (8 uL). Handling procedure 5 of Ferrozine method Iron Assay kit LS
(6)Incubate at room temperature for 5 minutes. Handling procedure 6 of Ferrozine method Iron Assay kit LS
(7)Read the absorbance at 560 nm (main) as OD2. Handling procedure 7 of Ferrozine method Iron Assay kit LS
(8)Calculate concentration.

    ΔODStd = (OD2Std - OD1Std) - (OD2Bl1 - ODBl)
    ΔODS = (OD2S - OD1S) - (OD2Bl - OD1Bl)
    Iron (ug/dL) = ΔODS/ΔODStd x 200
    Iron (uM) = ΔODS/ΔODStd x 35.8


  *ODS : OD sample, ODBl : OD blank,
   ODStd : OD Standard


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Citations

  1. Regulation of iron homeostasis by the p53-ISCU pathway; Sci Rep. 2015; 5: 16497
  2. Characterization of Flavin-Containing Opine Dehydrogenase from Bacteria, Plos One ; September 18, 2015
  3. Association between renal iron accumulation and renal interstitial fibrosis in a rat model of chronic kidney disease, Hypertension Research ; 38, 463-470 (July 2015)
  4. Temporary Dietary Iron Restriction Affects the Process of Thrombus Resolution in a Rat Model of Deep Vein Thrombosis, Plos One ; May 11, 2015
  5. JAK2V617F-positive myeloproliferative neoplasm clones evoke paracrine DNA damage to adjacent normal cells through secretion of lipocalin-2, Blood ; September 12, 2014
  6. Angiotensin II alters the expression of duodenal iron transporters, hepatic hepcidin, and body iron distribution in mice, Eur J Nutr ; August 7, 2014
  7. Iron Chelation by Deferoxamine Prevents Renal Interstitial Fibrosis in Mice with Unilateral Ureteral Obstruction, PLoS One ; February 19, 2014
  8. TEMPO-oxidized cellulose hydrogel as a high-capacity and reusable heavy metal ion adsorbent, J Hazard Mater ; September 15, 2013
  9. Superparamagnetic nanoparticle clusters for cancer theranostics combining magnetic resonance imaging and hyperthermia treatment, Theranostics ; April 23, 2013
  10. Dietary iron restriction inhibits progression of diabetic nephropathy in db/db mice, Am J Physiol Renal Physiol ; April 1, 2013
  11. GRK6 deficiency in mice causes autoimmune disease due to impaired apoptotic cell clearance, Nat Commun ; February 26, 2013
  12. Estrogen Regulates Hepcidin Expression via GPR30-BMP6-Dependent Signaling in Hepatocytes, PLoS One ; July 11, 2012
  13. FecA1, a bacterial iron transporter, determines the survival of Helicobacter pylori in the stomach, Free Radic Biol Med ; March 15, 2012

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Product Information

[Date : March 26 2025 00:07]

Detail Product Name Product Code Supplier Size Price
Iron Assay kit LS - Ferrozine method
DatasheetThis may not be the latest data sheet.
FE31ME AKJMetallogenics Co.,Ltd. 1 kit $296

Description Metallo Assay Iron Assay kit LS is intended for the quantitative determination of iron (: free iron , non-heme iron) in biological sample by 96-well reader use. Target: free iron (Fe2+ and Fe3+) Measuring range: 5-1,000 μg/Dl Species: all Sample: serum, plasma, urine, saliva, tissue extract, cell lysate, biological fluid, food, beverage etc Apparatus: UV/Vis 96-well reader or spectrophotometer Wavelength: 560 nm (540-580nm) Procedure: 10 min
Storage 4°C CAS
Link

Assay Kit Product List
Metallo Assay LS
"Metallo Assay Kit LS" in a minute!

[Date : March 26 2025 00:07]

Iron Assay kit LS - Ferrozine method


  • Product Code: FE31ME
  • Supplier: AKJ
  • Size: 1kit
  • Price: $296

Description Metallo Assay Iron Assay kit LS is intended for the quantitative determination of iron (: free iron , non-heme iron) in biological sample by 96-well reader use. Target: free iron (Fe2+ and Fe3+) Measuring range: 5-1,000 μg/Dl Species: all Sample: serum, plasma, urine, saliva, tissue extract, cell lysate, biological fluid, food, beverage etc Apparatus: UV/Vis 96-well reader or spectrophotometer Wavelength: 560 nm (540-580nm) Procedure: 10 min
Storage 4°C CAS
Link

Assay Kit Product List
Metallo Assay LS
"Metallo Assay Kit LS" in a minute!

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