Hepcidin ELISA Kit (Hepc)

Full Name: Hepcidin ELISA Kit (Hepc)
Reactivity: Human
Sample Type: Biological Fluids, Plasma, Tissue Homogenates, Serum
Sensitivity: 18.75 pg/ml

INTRODUCTION

Hepcidin is a tightly folded polypeptide containing 25 residues in length and is 32% beta sheets. The 25 amino acids have a hairpin structure and are stabilized by 4 disulfide bonds. It exists as a preprohormone (84 amino acids), prohormone (60 amino acids) and hormone (25 amino acids). Twenty- and 22-amino acid metabolites also exist in urine. The erythropoietin gene loses function and expresses more when the last five amino acids are removed. Oxygen-carrying hemoglobin binds to hepcidin, which affects oxygen binding and release. They are a family of glycoproteins that regulate iron absorption, transport and storage by binding to ferritin. On chromosome 3, the HFE gene, which encodes them, can be found. In order to detect whether there is too much or not enough of this protein, hepcidin levels can be assessed using blood serum testing or urine samples.

Hepcidins bind to receptors on the surface of cells called hepcidin-binding receptors (HBRs). There are six known HBRs, which all act as receptors. The hepcidin-binding receptors 2, 3 and 6 (HBRS2, HBRS3 and HBRS6) are responsible for regulating iron storage in the body. It binds to these three receptors to trigger an increase in the release of iron from stores in ferritin and transferrin. In addition, HBRS 1 is a known iron-regulating receptor. By turning on the erythropoietin receptor, which in turn increases erythropoiesis and red blood cell synthesis, it controls the iron utilisation route. It binds to this receptor to generate a cascade of signals that end in erythropoesis stimulation. This can also promote bone formation as well as intestinal absorption of iron, although this last process is more likely mediated by HBRS2.

Hepcidin is a hormone or peptide that regulates iron homeostasis in the body. It controls how much iron is available for functions like making hemoglobin and red blood cells and how much iron is stored in cells bound to ferritin. It also prevents iron overload and limits bacterial growth by reducing iron in plasma and extracellular fluids. Its levels can be affected by various iron disorders such as; hemochromatosis, anemia and hepatitis C. It is synthesized by hepatocytes, and its level increases when the body has a higher need for iron.

INTENDED USE

Human hepcidin ELISA kit can be used to measure levels of hepc (hepcidin, HAMP, PLTR, LEAP1, hepcidin-25) present is human biological fluids, serum, plasma and tissue homogenate samples.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

One 96-Well Plate: Pre-coated with anti-hepc antibody.
Standards: Lyophilized recombinant.
Sample/Standard Dilution Buffer.
Biotinylated-labelled Antibody.
Antibody Dilution Buffer.
HRP-Streptavidin Conjugate (SABC).
SABC Dilution Buffer.
TMB Substrate.
Wash Buffer (25x).
Plate Sealer.
Product Instructions.

TYPICAL RESULTS

For this hepcidin ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 31.25, 62.5, 125, 250, 500, 1000, 2000 pg/ml.
Reactivity: Human
Sensitivity: 18.75 pg/ml
Range: 31.25 – 2000 pg/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for hepc, no cross reactivity or interference between hepc and analogues was detected.
– Recovery: Serum (89 – 100%), EDTA Plasma (88 – 100%), Heparin Plasma (88 – 100%).
– Linearity: Serum (88 – 100%), EDTA Plasma (88 – 100%), Heparin Plasma (85 – 100%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

Hepcidin-Ferroportin Interaction Controls Systemic Iron Homeostasis. Int J Mol Sci. (2021) 22 (12): 6493. Nemeth E. and Ganz T.
Iron metabolism and iron disorders revisited in the hepcidin era. (2020) 105 (2): 260-272. Camaschella C., et al.
Structure of hepcidin-bound ferroportin reveals iron homeostatic mechanisms. (2020) 586 (7831): 807-811. Billesbølle C.B., et al.
Hepcidin and its therapeutic potential in neurodegenerative disorders. Med Res Rev. (2020) 40 (2): 633-653. Qian Z.M. and Ke Y.

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