Hepcidin 25 bioactive
The Hepcidin-25 (bioactive) ELISA is an enzyme immunoassay for measurement of Hepcidin-25 in serum and plasma.
Hepcidin is an iron homoeostasis regulator peptide. The bioactive peptide Hepcidin-25 is generated predominantly in the liver by proteolytic cleavage of the C-terminal 25 amino acids of prohepcidin (1). Subsequent N-terminal processing of Hepcidin-25 results in smaller peptides of 20-24 amino acids that show greatly reduced activity and accumulate in the urine (2). Although originally identified as antimicrobial peptide (3), Hepcidin-25 is now established as a major regulator of dietary iron absorption and cellular iron release (4). Hepcidin exerts its regulatory function by counteracting the function of ferroportin, the major cellular iron exporter in the membrane of macrophages, hepatocytes, and the basolateral site of enterocytes. Hepcidin-25 induces the internalization and degradation of ferroportin, resulting in increased intracellular iron stores, decreased dietary iron absorption, and decreased circulating iron concentrations (5). Hepatocellular hepcidin synthesis decreases under conditions of increased demand for circulating iron like iron deficiency, hypoxia, anemia, and erythropoiesis. In contrast, hepcidin synthesis is induced by inflammation and infection (6).
The Hepcidin-25 ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA), based on the principle of competitive binding. The microtiter wells are coated with a monoclonal (mouse) antibody directed towards an antigenic site of the Hepcidin-25 molecule. Endogenous Hepcidin-25 of a sample competes with a Hepcidin-25-biotin conjugate for binding to the coated antibody. After incubation, the unbound conjugate is washed off and a streptavidin-peroxidase enzyme complex is added to each well. After incubation, unbound enzyme complex is washed off and substrate solution is added. The blue colour development is stopped after a short incubation time, turning the colour from blue to yellow. The intensity of colour developed is reverse proportional to the concentration of Hepcidin in the sample.
- Scamuffa N, et al. Regulation of prohepcidin processing and activity by the subtilisin-like proprotein convertases Furin, PC5, PACE4 and PC7. Gut. 2008; 57(11):1573-82.
- Kemna EH, Tjalsma H, Podust VN, Swinkels DW. Mass spectrometry-based hepcidin measurements in serum and urine: analytical aspects and clinical implications. Clin. Chem. 2007; 53(4):620-8.
- Krause A et al. LEAP-1, a novel highly disulfide-bonded human peptide, exhibits antimicrobial activity. FEBS Lett. 2000; 480(2-3):147-50.
- Kroot J et al. Hepcidin in Human Iron Disorders: Diagnostic Implications. Clin. Chem. 2011; 57:121650–1669.
- Nemeth E, Ganz T. The role of hepcidin in iron metabolism. Acta Haematol. 2009; 122(2-3):78-86.
- Hentze MW, Muckenthaler MU, Galy B, Camaschella C. Two to tango: regulation of Mammalian iron metabolism. Cell. 2010; 142:24 –38.
- Swinkels DW, Jorna AT, Raymakers RA. Synopsis of the Dutch multidisciplinary guideline for the diagnosis and treatment of hereditary haemochromatosis. Neth. J. Med. 2007; 65(11):452-5.
- Camaschella C, Silvestri L. Molecular mechanisms regulating hepcidin revealed by hepcidin disorders. ScientificWorldJournal. 2011; 11:1357-66.
- Girelli D et al. Reduced serum hepcidin levels in patients with chronic hepatitis C. J. Hepatol. 2009; 51(5):845-52.
- Gardenghi S et al. Ineffective erythropoiesis in beta-thalassemia is characterized by increased iron absorption mediated by downregulation of hepcidin and up-regulation of ferroportin. Blood. 2007; 109:5027–35.
- Uehata T et al. Serum hepcidin-25 levels and anemia in non-dialysis chronic kidney disease patients: a cross-sectional study. Nephrol Dial. Transplant. 2012; 27(3):1076-83.
- Peeling P, Dawson B, Goodman C, Landers G, Trinder D. Athletic induced iron deficiency: new insights into the role of inflammation, cytokines and hormones. Eur. J. Appl. Physiol. 2008; 103: 381–91.