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ELISA Kit for Hepcidin (Hepc)

HAMP; HFE2B; PLTR; LEAP1; Hepcidin Antimicrobial Peptide; Liver-expressed antimicrobial peptide 1; Putative liver tumor regressor

Specificity

This assay has high sensitivity and excellent specificity for detection of Hepcidin (Hepc).
No significant cross-reactivity or interference between Hepcidin (Hepc) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Hepcidin (Hepc) and the recovery rates were calculated by comparing the measured value to the expected amount of Hepcidin (Hepc) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-90 86
EDTA plasma(n=5) 91-98 95
heparin plasma(n=5) 98-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hepcidin (Hepc) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Hepcidin (Hepc) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Hepcidin (Hepc) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 86-94% 83-102% 97-105% 94-102%
EDTA plasma(n=5) 88-98% 79-95% 84-104% 78-96%
heparin plasma(n=5) 92-101% 86-101% 98-105% 80-102%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
World J Gastroenterol. Hepcidin expression in colon during trinitrobenzene sulfonic acid-induced colitis in rats Pubmed: 24764672
International Journal of Advanced Research Modulations of serum hepcidin, IL6 and iron status by different exercise regimens in ovarectomized rats Com:Source
Neurochemical Journal Serum iron metabolism markers including hepcidin in multiple sclerosis patients Springer:Source
Afro-Egypt J Infect Endem Dis Monitoring Hepcidin Level in Chronic Hepatitis C Virus Patients during Therapy Zu:Source
Gastroenterology Gluconeogenic signals regulate iron homeostasis via hepcidin in mice Pubmed:Pmc3989026
J Card Fail. Inappropriate expression of hepcidin by liver congestion contributes to anemia and relative iron deficiency Pubmed:24440572
Food Funct. The action of JAK, SMAD and ERK signal pathways on hepcidin suppression by polysaccharides from Angelica sinensis in rats with iron deficiency anemia Pubmed:24752529
Free Radic Res. Changes in skeletal muscle iron metabolism outpace amyotrophic lateral sclerosis onset in transgenic rats bearing the G93A hmSOD1 gene mutation Pubmed:25175826
Saudi J Kidney Dis Transpl. Hepcidin and regulation of iron homeostasis in maintenance hemodialysis patients Pubmed:25193892
J Endocrinol Invest.? The interplay among iron metabolism, endothelium and inflammatory cascade in dysmetabolic disorders Pubmed:25245337
Gastroenterology Research and Practice An Evaluation of the Correlation between Hepcidin Serum Levels and Disease Activity in Inflammatory Bowel Disease Pubmed:25628652
Clin Lab. Serum hepcidin levels in Bulgarian population. Pubmed:25651734
The Tohoku Journal of Experimental Medicine Liver Congestion in Heart Failure Contributes to Inappropriately Increased Serum Hepcidin despite Anemia Pubmed:25742771
Cancer Res Hepcidin regulation in prostate and its disruption in prostate cancer PubMed: 25858146
Biochem Biophys Res Commun B-cell translocation gene 2 promotes hepatic hepcidin production via induction of Yin Yang 1 PubMed: 25839654
Blood Purif Effect of a More Permeable Dialysis Membrane on ESA Resistance in Hemodialysis Patients - A Pilot Investigation PubMed: 26536083
Asian Journal of Clinical Nutrition Urinary Hepcidin in Diagnosis of Iron Deficiency Anemia Openview: E01Ad7C4A9109Ed5Fadb03Ec206D7Bdc
Front Psychol Ferrous sulfate, but not iron polymaltose complex, aggravates local and systemic inflammation and oxidative stress in dextran sodium sulfate-induced colitis in rats PubMed: 25852597
Biol Trace Elem Res Iron Supplementation Attenuates the Inflammatory Status of Anemic Piglets by Regulating Hepcidin PubMed: 25774043
APS Journals Kupffer cell depletion protects against the steatosis, but not the liver damage, induced by marginal copper, high fructose diet in male rats. PubMed: 25813056
European Journal of Haematology Hepcidin in tumor‐related iron deficiency anemia and tumor‐related anemia of chronic disease: pathogenic mechanisms and diagnosis PubMed: 24954786
WJA The Effects of Treatment on Serum Hepcidin and Iron Homeostasis in HIV-1-Infected In-dividuals Journal: 59107
Nutrition, Metabolism and Cardiovascular Diseases Hepcidin and iron metabolism associated with cardiometabolic risk factors in children: A case-control study. Pubmed:27139516
Food Funct The effects of polysaccharides from the root of Angelica sinensis on tumor growth and iron metabolism in H22-bearing mice Pubmed:26757699
Antioxid Redox Signal. Hydrogen sulfide attenuates inflammatory hepcidin by reducing IL-6 secretion and promoting SIRT1-mediated STAT3 deacetylation Pubmed:26154696
BioMed Research International The Impact of Acute Matriptase Inhibition in Hepatic Inflammatory Models. pubmed:27642598
Frontiers in Neuroscience In Absence of the Cellular Prion Protein, Alterations in Copper Metabolism and Copper-Dependent Oxidase Activity Affect Iron Distribution. pubmed:27729845
Experimental Biology and Medicine Effects of iron supplementation in mice with hypoferremia induced by obesity pubmed:27439539
Nutrition, Metabolism and Cardiovascular Diseases Hepcidin and iron metabolism associated with cardiometabolic risk factors in children: A case–control study pubmed:27139516
PLOS ONE S-Propargyl-Cysteine, a Novel Hydrogen Sulfide Donor, Inhibits Inflammatory Hepcidin and Relieves Anemia of Inflammation by Inhibiting IL-6/STAT3 Pathway. pubmed:27649298
Endocrine connections Hepcidin links gluco-toxicity to pancreatic beta cell dysfunction by inhibiting Pdx-1 expression pubmed:28179377
Maternal and cord blood hepcidin levels based on gestational weeks in term and preterm infants publication:305921433_Maternal_and_cord_blood_hepc
Oxidative Medicine and Cellular Longevity Acidic Polysaccharide from Angelica sinensis Reverses Anemia of Chronic Disease Involving the Suppression of Inflammatory Hepcidin and NF-κB Activation pubmed:29147463
The Royal Society of Chemistry Silkworm feces extract improves iron deficiency anemia via suppressing hepcidin expression and promoting iron-regulatory proteins expression DOI: 10.1039/C7RA09576G
Journal of Inflammation Evaluation of a competitive hepcidin ELISA assay in the differential diagnosis of iron deficiency anaemia with concurrent inflammation and anaemia of inflammation in elderly patients 10.1186:s12950-017-0166-3
Antioxidants and Redox Signaling AMPK serves as a Therapeutic Target against Anemia of Inflammation pubmed:27923278
NutriciónHospitalaria Micronutrient supplementation in gastric bypass surgery: prospective study on infl ammation and iron metabolism in premenopausal women 10.20960/nh.1162
The Japanese Society of Hematology Hepcidin suppression in β‑thalassemia is associated with the down‑regulation of atonal homolog 8 pubmed:28405918
Tuberculosis Modulation of iron status biomarkers in tuberculosis-diabetes co-morbidity 10.1016/j.tube.2017.11.011
Oman Medical Journal The Role of Insulin Therapy in Correcting Hepcidin Levels in Patients with Type 2 Diabetes Mellitus. pubmed:28584599
Cellular Physiology and Biochemistry The Effects of Angelica Sinensis Polysaccharide on Tumor Growth and Iron Metabolism by Regulating Hepcidin in Tumor-Bearing Mice Pubmed:29843136
Cellular Microbiology  inhibits ferroportin translation by modulating FBXL5‐IRP2 axis for its growth within host macrophages Pubmed:29470856
Journal of Histochemistry & Cytochemistry Acute and Chronic Iron Overloading Differentially Modulates the Expression of Cellular Iron-homeostatic Molecules in Normal Rat Kidney Pubmed:29873589
Clinical and Experimental Medicine Increased NGAL level associated with iron store in chronic kidney disease with anemia Pubmed:29909502
Hematology Relationship between obesity and iron deficiency anemia: is there a role of hepcidin? Pubmed:29318945
Hepcidin serum levels in HCV chronically mono-infected naïve patients, compared to healthy individuals 77856
Food & Function Effect of iron supplement from microalgae on the treatment of iron-deficient anemia in rats Doi: 10.1039/C8FO01696H
Haematologica New thiazolidinones reduce iron overload in mouse models of hereditary hemochromatosis and β-thalassemia Pubmed: 30792208
Ann Intensive Care Inflammatory anemia-associated parameters are related to 28-day mortality in patients with sepsis admitted to the ICU: a preliminary observational study Pubmed: 31183575
Journal of Functional Foods Dietary gelatin enhances non-heme iron absorption possibly via regulation of systemic iron homeostasis in rats
Naunyn Schmiedebergs Arch Pharmacol. 3-Amidinophenylalanine-derived matriptase inhibitors can modulate hepcidin production in vitro Pubmed: 31659405
DEVELOPMENT GROWTH & DIFFERENTIATION H2O2 induces oxidative stress damage through the BMP‐6/SMAD/hepcidin axis Pubmed: 32012242
Frontiers in Physiology Increased Hepcidin Levels During a Period of High Training Load Do Not Alter Iron Status in Male Elite Junior Rowers Pubmed: 32038278
J Korean Med Sci Can Calprotectin Show Subclinical Inflammation in Familial Mediterranean Fever Patients? Pubmed: 32174062
nephrology dialysis transplantation Iron kinetics following treatment with sucroferric oxyhydroxide or ferric citrate in healthy rats and models of anaemia, iron overload or inflammation Pubmed: 32259248
mikrochimica acta Gold-silver core-shell nanoparticle–based impedimetric immunosensor for detection of iron homeostasis biomarker hepcidin Pubmed: 33095336
JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY Circulating Osteocyte‐Related Biomarkers (vitamin D, sclerostin, dickkopf-1), hepcidin, and oxidative stress markers in early breast cancer: their impact in … Pubmed: 33065276
Sleep Breath Relation of serum hepcidin levels and restless legs syndrome in chronic hemodialysis patients Pubmed: 33029690
journal of trace elements in experimental medicine Difference in iron metabolism may partly explain sex-related variability in the manifestation of Wilson's disease Pubmed: 32937238
reproduction in domestic animals Expression of iron regulatory proteins in full term swine placenta Pubmed: 32449967
Neurobiology of Aging Estrogen deficiency is associated with brain iron deposition via upregulation of hepcidin expression in aged female mice Pubmed: 32920472
The Egyptian Journal of Haematology Urinary hepcidin concentration in assessment of iron homeostasis in pediatrics
COMPARATIVE MONITORING OF THE ENZYME ACTIVITY OF BONE METABOLISM IN PATIENTS AFTER DENTAL IMPLANTATION
Serum iron, prealbumin, D-dimer and prothrombin time are indicators of the diagnosis, severity and outcome of Salmonella enterica serovar Enteritidis infection
Гепсидин и индекс фиброза печени MELD-XI как маркеры полиорганной недостаточности у пациентов хронической сердечной недостаточностью с …
Ассоциации уровней NT-proBNP и гепсидина с клинико-лабораторными параметрами у больных хронической сердечной недостаточностью с разной …
Biosci Rep Correlation of hepcidin and serum ferritin levels in thalassemia patients at Chiang Mai University Hospital 33565577
Biometals Iron metabolism is disturbed and anti-copper treatment improves but does not normalize iron metabolism in Wilson's disease 33555495
J Mol Cell Cardiol Systemic iron deficiency does not affect the cardiac iron content and progression of heart failure 34139233
Continuous Veno-Venous Hemofiltration (CVVH) Improves Iron Metabolism Disorders in Patients With Sepsis: A Single-Center Prospective Cohort Study
Reports of Biochemistry & Molecular Biology Evaluation of Serum Cyclooxygenase, Hepcidin Levels in Acute Renal Injury (AKI) Patients Following Cardiac Catheterization
Food Funct High-dose ferric citrate supplementation attenuates omega-3 polyunsaturated fatty acid biosynthesis via downregulating delta 5 and 6 desaturases in rats with high-fat … 34787162
J Trace Elem Med Biol Caffeine and alcohol-friends or foes of human iron stores? Pubmed:35063815
PeerJ Resveratrol ameliorates iron overload induced liver fibrosis in mice by regulating iron homeostasis Pubmed:35698613
Serum Calprotectin and Iron Metabolism Biomarker Levels in Behçet's Disease
Food Bioscience Roles of homopolymeric apoferritin in alleviating alcohol-induced liver injury
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