Nitroso-PSAP method Iron Concentration Assay Kit Iron Assay kit LS | Metallo Assay LS

• Features
• Assay Principle
• Sample Preparation
• Handling Procedure
• Citations
• Product Information

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.
• Assay range: 10-1,000 μg/dL
• 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 (r^2=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 (#UIB01E), TIBC (Total Iron Binding Capacity) can be measured.

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Assay Principle

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Sample Preparation

• 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. transferrin, ferritin, 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 (15 uL) to 96 well plate.
(2)Add R-A buffer (160 uL).
(3)Incubate at room temperature for 10 minutes.
(4)Add working reagent (75 uL).
(5)Incubate at room temperature for 5 minutes.
(6)Read the absorbance at 750 nm (main).
(7)Calculate concentration.     ΔODStd = ODStd - ODBl, ΔODS = ODS - ODBl     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

No.	Literature information	Remarks	URL
1	Kagoya Y et al. JAK2V617F+ myeloproliferative neoplasm clones evoke paracrine DNA damage to adjacent normal cells through secretion of lipocalin-2. Blood 2014 Nov;124(19):2996-3006 Kagoya Y et al 2014/01/01
2	Tajima S et al. Angiotensin II alters the expression of duodenal iron transporters, hepatic hepcidin, and body iron distribution in mice. Eur J Nutr 2014 Aug; Tajima S et al 2014/01/01
3	Hayashi K et al. Superparamagnetic nanoparticle clusters for cancer theranostics combining magnetic resonance imaging and hyperthermia treatment. Theranostics 2013;3(6):366-76 Hayashi K et al 2013/01/01
4	Tsugawa H et al. FecA1, a bacterial iron transporter, determines the survival of Helicobacter pylori in the stomach. Free Radic. Biol. Med. 2012 Mar;52(6):1003-10 Tsugawa H et al 2012/01/01
5	Involvement of splenic iron accumulation in the development of nonalcoholic steatohepatitis in Tsumura Suzuki Obese Diabetes mice. Scientific Reports 6; Article number: 22476, 02 March 2016
6	Iron-induced skeletal muscle atrophy involves an Akt-forkhead box O3–E3 ubiquitin ligase-dependent pathway Biology; Volume 35, May 2016, Pages 66–76
7	Characterization of Flavin-Containing Opine Dehydrogenase from Bacteria　Plos One; September 18, 2015
8	Association between renal iron accumulation and renal interstitial fibrosis in a rat model of chronic kidney disease　Hypertension Research;38, 463-470 (July 2015)
9	Temporary Dietary Iron Restriction Affects the Process of Thrombus Resolution in a Rat Model of Deep Vein Thrombosis　Plos one; May 11, 2015
10	Involvement of thioredoxin on the scaffold activity of NifU in heterocyst cells of the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120　J Biochem first published online; May 6, 2015
11	Sexual development of Schizosaccharomyces pombe is induced by zinc or iron limitation through Ecl1 family genes. Molecular Genetics and Genomics, February 2015, Volume 290, Issue 1, pp 173-185
12	Association of dietary iron restriction with left ventricular remodeling after myocardial infarction in mice　Heart and Vessels; 09 January 2015
13	Oshima K et al. Iron suppresses erythropoietin expression via oxidative stress-dependent hypoxia-inducible factor-2 alpha inactivation. Lab. Invest. 2017 May;97(5):555-566 Oshima K et al 2017/01/01
14	Yoshida K et al. Distinct electron transfer from ferredoxin-thioredoxin reductase to multiple thioredoxin isoforms in chloroplasts. Biochem. J. 2017 Apr;474(8):1347-1360 Yoshida K et al 2017/01/01
15	Tsujino S et al. Pyruvic oxime dioxygenase from heterotrophic nitrifier Alcaligenes faecalis is a nonheme Fe((II))-dependent enzyme homologous to class II aldolase. Sci Rep 2017 Jan;7:39991 Tsujino S et al 2017/01/01
16	Watanabe S et al. Functional characterization of aconitase X as a cis-3-hydroxy-L-proline dehydratase. Sci Rep 2016 Dec;6:38720 Watanabe S et al 2016/01/01
17	Murotomi K et al. Involvement of splenic iron accumulation in the development of nonalcoholic steatohepatitis in Tsumura Suzuki Obese Diabetes mice. Sci Rep 2016 Mar;6:22476 Murotomi K et al 2016/01/01
18	Ikeda Y et al. Iron-induced skeletal muscle atrophy involves an Akt-forkhead box O3-E3 ubiquitin ligase-dependent pathway. J Trace Elem Med Biol 2016 May;35:66-76 Ikeda Y et al 2016/01/01
19	Watanabe S et al. Characterization of Flavin-Containing Opine Dehydrogenase from Bacteria. PLoS ONE 2015;10(9):e0138434 Watanabe S et al 2015/01/01
20	Naito Y et al. Association between renal iron accumulation and renal interstitial fibrosis in a rat model of chronic kidney disease. Hypertens. Res. 2015 Jul;38(7):463-70 Naito Y et al 2015/01/01
21	Oboshi M et al. Temporary dietary iron restriction affects the process of thrombus resolution in a rat model of deep vein thrombosis. PLoS ONE 2015;10(5):e0126611 Oboshi M et al 2015/01/01
22	Nomata J et al. Involvement of thioredoxin on the scaffold activity of NifU in heterocyst cells of the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120. J. Biochem. 2015 Sep;158(3):253-61 Nomata J et al 2015/01/01
23	Ohtsuka H et al. Sexual development of Schizosaccharomyces pombe is induced by zinc or iron limitation through Ecl1 family genes. Mol. Genet. Genomics 2015 Feb;290(1):173-85 Ohtsuka H et al 2015/01/01
24	Eguchi A et al. Association of dietary iron restriction with left ventricular remodeling after myocardial infarction in mice. Heart Vessels 2016 Feb;31(2):222-9 Eguchi A et al 2016/01/01
25	Imamura T et al. Hydroxyl radicals cause fluctuation in intracellular ferrous ion levels upon light exposure during photoreceptor cell death. Exp. Eye Res. 2014 Dec;129:24-30 Imamura T et al 2014/01/01
26	Ikeda Y et al. Iron chelation by deferoxamine prevents renal interstitial fibrosis in mice with unilateral ureteral obstruction. PLoS ONE 2014;9(2):e89355 Ikeda Y et al 2014/01/01
27	Isobe N et al. TEMPO-oxidized cellulose hydrogel as a high-capacity and reusable heavy metal ion adsorbent. J. Hazard. Mater. 2013 Sep;260:195-201 Isobe N et al 2013/01/01
28	Ikeda Y et al. Dietary iron restriction inhibits progression of diabetic nephropathy in db/db mice. Am. J. Physiol. Renal Physiol. 2013 Apr;304(7):F1028-36 Ikeda Y et al 2013/01/01
29	Nakaya M et al. GRK6 deficiency in mice causes autoimmune disease due to impaired apoptotic cell clearance. Nat Commun 2013;4:1532 Nakaya M et al 2013/01/01
30	Ikeda Y et al. Estrogen regulates hepcidin expression via GPR30-BMP6-dependent signaling in hepatocytes. PLoS ONE 2012;7(7):e40465 Ikeda Y et al 2012/01/01

• No.： 1
• Literature information：
  Kagoya Y et al. JAK2V617F+ myeloproliferative neoplasm clones evoke paracrine DNA damage to adjacent normal cells through secretion of lipocalin-2. Blood 2014 Nov;124(19):2996-3006
  Kagoya Y et al
  2014/01/01
  
• Remarks：
  
• URL：
  

• No.： 2
• Literature information：
  Tajima S et al. Angiotensin II alters the expression of duodenal iron transporters, hepatic hepcidin, and body iron distribution in mice. Eur J Nutr 2014 Aug;
  Tajima S et al
  2014/01/01
  
• Remarks：
  
• URL：
  

• No.： 3
• Literature information：
  Hayashi K et al. Superparamagnetic nanoparticle clusters for cancer theranostics combining magnetic resonance imaging and hyperthermia treatment. Theranostics 2013;3(6):366-76
  Hayashi K et al
  2013/01/01
  
• Remarks：
  
• URL：
  

• No.： 4
• Literature information：
  Tsugawa H et al. FecA1, a bacterial iron transporter, determines the survival of Helicobacter pylori in the stomach. Free Radic. Biol. Med. 2012 Mar;52(6):1003-10
  Tsugawa H et al
  2012/01/01
  
• Remarks：
  
• URL：
  

• No.： 5
• Literature information：
  Involvement of splenic iron accumulation in the development of nonalcoholic steatohepatitis in Tsumura Suzuki Obese Diabetes mice. Scientific Reports 6; Article number: 22476, 02 March 2016
  
  
• Remarks：
  
• URL：
  

• No.： 6
• Literature information：
  Iron-induced skeletal muscle atrophy involves an Akt-forkhead box O3–E3 ubiquitin ligase-dependent pathway Biology; Volume 35, May 2016, Pages 66–76
  
  
• Remarks：
  
• URL：
  

• No.： 7
• Literature information：
  Characterization of Flavin-Containing Opine Dehydrogenase from Bacteria　Plos One; September 18, 2015
  
  
• Remarks：
  
• URL：
  

• No.： 8
• Literature information：
  Association between renal iron accumulation and renal interstitial fibrosis in a rat model of chronic kidney disease　Hypertension Research;38, 463-470 (July 2015)
  
  
• Remarks：
  
• URL：
  

• No.： 9
• Literature information：
  Temporary Dietary Iron Restriction Affects the Process of Thrombus Resolution in a Rat Model of Deep Vein Thrombosis　Plos one; May 11, 2015
  
  
• Remarks：
  
• URL：
  

• No.： 10
• Literature information：
  Involvement of thioredoxin on the scaffold activity of NifU in heterocyst cells of the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120　J Biochem first published online; May 6, 2015
  
  
• Remarks：
  
• URL：
  

• No.： 11
• Literature information：
  Sexual development of Schizosaccharomyces pombe is induced by zinc or iron limitation through Ecl1 family genes. Molecular Genetics and Genomics, February 2015, Volume 290, Issue 1, pp 173-185
  
  
• Remarks：
  
• URL：
  

• No.： 12
• Literature information：
  Association of dietary iron restriction with left ventricular remodeling after myocardial infarction in mice　Heart and Vessels; 09 January 2015
  
  
• Remarks：
  
• URL：
  

• No.： 13
• Literature information：
  Oshima K et al. Iron suppresses erythropoietin expression via oxidative stress-dependent hypoxia-inducible factor-2 alpha inactivation. Lab. Invest. 2017 May;97(5):555-566
  Oshima K et al
  2017/01/01
  
• Remarks：
  
• URL：
  

• No.： 14
• Literature information：
  Yoshida K et al. Distinct electron transfer from ferredoxin-thioredoxin reductase to multiple thioredoxin isoforms in chloroplasts. Biochem. J. 2017 Apr;474(8):1347-1360
  Yoshida K et al
  2017/01/01
  
• Remarks：
  
• URL：
  

• No.： 15
• Literature information：
  Tsujino S et al. Pyruvic oxime dioxygenase from heterotrophic nitrifier Alcaligenes faecalis is a nonheme Fe((II))-dependent enzyme homologous to class II aldolase. Sci Rep 2017 Jan;7:39991
  Tsujino S et al
  2017/01/01
  
• Remarks：
  
• URL：
  

• No.： 16
• Literature information：
  Watanabe S et al. Functional characterization of aconitase X as a cis-3-hydroxy-L-proline dehydratase. Sci Rep 2016 Dec;6:38720
  Watanabe S et al
  2016/01/01
  
• Remarks：
  
• URL：
  

• No.： 17
• Literature information：
  Murotomi K et al. Involvement of splenic iron accumulation in the development of nonalcoholic steatohepatitis in Tsumura Suzuki Obese Diabetes mice. Sci Rep 2016 Mar;6:22476
  Murotomi K et al
  2016/01/01
  
• Remarks：
  
• URL：
  

• No.： 18
• Literature information：
  Ikeda Y et al. Iron-induced skeletal muscle atrophy involves an Akt-forkhead box O3-E3 ubiquitin ligase-dependent pathway. J Trace Elem Med Biol 2016 May;35:66-76
  Ikeda Y et al
  2016/01/01
  
• Remarks：
  
• URL：
  

• No.： 19
• Literature information：
  Watanabe S et al. Characterization of Flavin-Containing Opine Dehydrogenase from Bacteria. PLoS ONE 2015;10(9):e0138434
  Watanabe S et al
  2015/01/01
  
• Remarks：
  
• URL：
  

• No.： 20
• Literature information：
  Naito Y et al. Association between renal iron accumulation and renal interstitial fibrosis in a rat model of chronic kidney disease. Hypertens. Res. 2015 Jul;38(7):463-70
  Naito Y et al
  2015/01/01
  
• Remarks：
  
• URL：
  

• No.： 21
• Literature information：
  Oboshi M et al. Temporary dietary iron restriction affects the process of thrombus resolution in a rat model of deep vein thrombosis. PLoS ONE 2015;10(5):e0126611
  Oboshi M et al
  2015/01/01
  
• Remarks：
  
• URL：
  

• No.： 22
• Literature information：
  Nomata J et al. Involvement of thioredoxin on the scaffold activity of NifU in heterocyst cells of the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120. J. Biochem. 2015 Sep;158(3):253-61
  Nomata J et al
  2015/01/01
  
• Remarks：
  
• URL：
  

• No.： 23
• Literature information：
  Ohtsuka H et al. Sexual development of Schizosaccharomyces pombe is induced by zinc or iron limitation through Ecl1 family genes. Mol. Genet. Genomics 2015 Feb;290(1):173-85
  Ohtsuka H et al
  2015/01/01
  
• Remarks：
  
• URL：
  

• No.： 24
• Literature information：
  Eguchi A et al. Association of dietary iron restriction with left ventricular remodeling after myocardial infarction in mice. Heart Vessels 2016 Feb;31(2):222-9
  Eguchi A et al
  2016/01/01
  
• Remarks：
  
• URL：
  

• No.： 25
• Literature information：
  Imamura T et al. Hydroxyl radicals cause fluctuation in intracellular ferrous ion levels upon light exposure during photoreceptor cell death. Exp. Eye Res. 2014 Dec;129:24-30
  Imamura T et al
  2014/01/01
  
• Remarks：
  
• URL：
  

• No.： 26
• Literature information：
  Ikeda Y et al. Iron chelation by deferoxamine prevents renal interstitial fibrosis in mice with unilateral ureteral obstruction. PLoS ONE 2014;9(2):e89355
  Ikeda Y et al
  2014/01/01
  
• Remarks：
  
• URL：
  

• No.： 27
• Literature information：
  Isobe N et al. TEMPO-oxidized cellulose hydrogel as a high-capacity and reusable heavy metal ion adsorbent. J. Hazard. Mater. 2013 Sep;260:195-201
  Isobe N et al
  2013/01/01
  
• Remarks：
  
• URL：
  

• No.： 28
• Literature information：
  Ikeda Y et al. Dietary iron restriction inhibits progression of diabetic nephropathy in db/db mice. Am. J. Physiol. Renal Physiol. 2013 Apr;304(7):F1028-36
  Ikeda Y et al
  2013/01/01
  
• Remarks：
  
• URL：
  

• No.： 29
• Literature information：
  Nakaya M et al. GRK6 deficiency in mice causes autoimmune disease due to impaired apoptotic cell clearance. Nat Commun 2013;4:1532
  Nakaya M et al
  2013/01/01
  
• Remarks：
  
• URL：
  

• No.： 30
• Literature information：
  Ikeda Y et al. Estrogen regulates hepcidin expression via GPR30-BMP6-dependent signaling in hepatocytes. PLoS ONE 2012;7(7):e40465
  Ikeda Y et al
  2012/01/01
  
• Remarks：
  
• URL：
  

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Product Information

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