| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
MMP-9 cathepsin K
Ferroportin (FPN1). Hepcidin-1 (mouse) is a 25-amino acid peptide hormone that acts as the master regulator of systemic iron homeostasis. Its primary molecular target is ferroportin (FPN1), which is the only known cellular iron exporter expressed on the basolateral surface of enterocytes (duodenal cells), macrophages (recycling iron from senescent red blood cells), and hepatocytes. Hepcidin binds directly to ferroportin, inducing its internalization, ubiquitination, and lysosomal degradation. This inhibits the export of iron into the plasma, thereby reducing serum iron levels. Hepcidin expression is regulated by circulating iron levels, erythropoietic demand, and inflammatory signals. By upregulating hepcidin, the body limits iron availability to invading pathogens (a host defense mechanism). Dysregulation of hepcidin is implicated in various disorders, including hereditary hemochromatosis (low hepcidin leading to iron overload) and anemia of chronic disease (high hepcidin leading to iron restriction). In vitro, Hepcidin-1 (mouse) TFA upregulates mRNA levels of TRAP, cathepsin K, and MMP-9, and increases TRAP-5b protein secretion. It also reduces FPN1 protein expression, leading to increased intracellular iron levels. The TFA salt does not affect activity. |
|---|---|
| ln Vitro |
Hepcidin-1 (mouse) TFA (200–800 nM, 4 days) facilitates the differentiation of raw264 produced by RANKL (50 ng/mL).7 cells [1]. The expression of TRAP, CTK, and MMP-9 mRNA is upregulated by hepcidin-1 (mouse) TFA (0-800 nM, 4 days) [1]. Ferroportin 1 (FPN1) protein is decreased in RAW264 and Trap-5b protein levels are raised by Hepcidin-1 (mouse) TFA (0-800 nM, 20 hours).7 cells [1].
In vitro, Hepcidin-1 (mouse) TFA is a key regulator of iron metabolism. In cell-based assays using ferroportin-expressing cells (e.g., macrophages or enterocyte-like cells), treatment with hepcidin-1 (0.1-10 uM) leads to a time- and dose-dependent decrease in ferroportin (FPN1) protein levels, as assessed by Western blot. This results in reduced iron export from cells, which can be measured by using 59Fe-labeled transferrin or by assessing intracellular iron levels. Hepcidin-1 (mouse) TFA upregulates the mRNA levels of TRAP, cathepsin K, and MMP-9, and increases TRAP-5b protein secretion. In osteoclasts, it has been shown to promote their differentiation and activity, which may contribute to bone loss in iron overload conditions. The peptide also acts as an antimicrobial peptide (part of the innate immune response), directly killing certain bacteria and fungi. The TFA salt is used for solubility. The EC50 for ferroportin degradation is typically in the 10-100 nM range. In cell viability assays, hepcidin is not cytotoxic at concentrations up to 10 uM for 24-48 hours. |
| ln Vivo |
In vivo, Hepcidin-1 (mouse) TFA induces hypoferremia (low serum iron levels) in mouse models. Intraperitoneal (i.p.) or subcutaneous (s.c.) administration of the peptide at doses of 1-10 mg/kg leads to a rapid and dose-dependent decrease in serum iron levels within 1-4 hours, as measured by colorimetric assays. This effect is mediated by the degradation of ferroportin in enterocytes and macrophages, blocking iron efflux into the circulation. In mouse models of iron overload (e.g., Hfe-/- mice or iron-dextran-treated mice), repeated administration of hepcidin-1 (1-5 mg/kg, i.p., daily for 7-14 days) reduces liver and spleen iron content, as measured by atomic absorption spectroscopy or Perl's Prussian blue staining. It also improves parameters of anemia in models of anemia of chronic disease (e.g., turpentine-induced inflammation) by restricting iron availability for erythro-poiesis. In models of inflammation (e.g., LPS-induced sepsis), hepcidin-1 levels are already elevated; exogenous hepcidin may exacerbate anemia. Hepcidin-1 has also been studied in models of osteoporosis, where it promotes osteoclast activity and bone resorption. The TFA salt is suitable for in vivo administration. All animal procedures must be approved by IACUC.
|
| Enzyme Assay |
For non-cellular binding assays, a direct binding assay between hepcidin and ferroportin can be performed using surface plasmon resonance (SPR). Recombinant human or mouse ferroportin protein (purified and reconstituted in nanodiscs or detergent micelles) is immobilized on a CM5 sensor chip via amine coupling. Hepcidin-1 (mouse) TFA is dissolved in running buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.005% P20, 1 mM DTT) at concentrations ranging from 0.1-1000 nM. Flow over the immobilized ferroportin at 25degC at a flow rate of 30 uL/min. Association and dissociation phases are recorded, and the KD is calculated using a 1:1 Langmuir binding model. Alternatively, a competitive binding assay using biotinylated hepcidin and streptavidin-HRP detection can be performed on immobilized ferroportin in an ELISA format. For a functional degradation assay in a cell-free system, no such system exists; degradation requires intact cells. For a binding assay using surface-expressed ferroportin: Use HEK293 cells expressing N-terminally HA-tagged or FLAG-tagged ferroportin. Incubate cells with varying concentrations of biotinylated hepcidin-1 (biotin-hepcidin) for 30-60 min at 4degC. After washing, lyse cells, capture biotinylated hepcidin-streptavidin agarose, and quantify bound ferroportin by Western blot. The Kd is determined.
|
| Cell Assay |
Western Blot Analysis[1]
Cell Types: RAW264.7 cells Tested Concentrations: 0, 200, 400, or 800 nM Incubation Duration: 20 h Experimental Results: diminished Ferroportin 1(FPN1) protein. RT-PCR[1] Cell Types: RAW264. 7 cells Tested Concentrations: 0, 200, 400, or 800 nM Incubation Duration: 4 days Experimental Results: Increased the gene expression of TRAP, CTK, and MMP-9 in a dose-dependent manner. For cellular assays, use cells expressing ferroportin: J774 murine macrophages, RAW 264.7 cells, or HEK293 cells stably expressing ferroportin with a tag. Seed cells in 6- or 12-well plates (2-5 × 10^5 cells/well) in DMEM with 10% FBS and culture overnight at 37degC, 5% CO2. Treat cells with Hepcidin-1 (mouse) TFA at concentrations of 0.01-10 uM for 2-24 hours. For ferroportin degradation assays, lyse cells in RIPA buffer with protease inhibitors, separate lysates (20-40 ug protein) by SDS-PAGE, and immunoblot with anti-ferroportin antibody (or anti-tag antibody). Quantify band density by densitometry and normalize to loading control (beta-actin or GAPDH). For iron export assays, load cells with 59Fe-transferrin (0.5-1 uCi/mL) for 2-4 hours, wash, and then treat with hepcidin (0.1-10 uM) for 2-24 hours. Collect the medium and measure 59Fe counts in a gamma counter. Hepcidin should reduce 59Fe export in a dose-dependent manner. For intracellular iron measurements, use a ferrozine-based colorimetric assay or the fluorescent iron probe Calcein-AM (quenched by iron). Treatment with hepcidin increases intracellular iron. For osteoclast differentiation assays, use bone marrow-derived macrophages (BMMs) treated with M-CSF (30 ng/mL) and RANKL (50 ng/mL) for 5-7 days. Add hepcidin (0.1-10 uM) to the medium. Quantify TRAP (tartrate-resistant acid phosphatase) staining (a marker of osteoclasts) and TRAP activity by colorimetric assay. Hepcidin promotes osteoclast differentiation. For antibacterial assays, incubate bacteria (e.g., E. coli, S. aureus, or Candida albicans) with hepcidin (1-100 uM) in low-salt buffer for 2-4 hours. Plate on agar and count colony-forming units (CFU). Hepcidin has direct antimicrobial activity. The TFA salt is water-soluble. All experiments should include a scrambled hepcidin peptide as a negative control. |
| Animal Protocol |
For in vivo studies, use female C57BL/6J mice (8-12 weeks old, 18-22 g). For acute hypoferremia studies, dissolve Hepcidin-1 (mouse) TFA in sterile saline or PBS (0.5-1 mg/mL). Administer the peptide intraperitoneally (i.p.) at doses of 1, 2.5, 5, and 10 mg/kg (volume 10 mL/kg). Collect blood via tail vein at pre-dose, 1, 2, 4, 6, 12, and 24 hours post-dose. Separate serum by centrifugation (2,000 × g, 10 min, 4degC). Measure serum iron levels using a colorimetric iron assay kit (e.g., QuantiChrom Iron Assay Kit). Hepcidin should reduce serum iron levels by 30-70% within 2-4 hours, with recovery by 24 hours. For chronic studies (iron overload models), administer iron dextran (100 mg/kg, i.p.) twice weekly for 2 weeks to induce iron overload in C57BL/6J mice. Then, treat with hepcidin-1 (1-5 mg/kg, i.p.) daily for 10-14 days. Vehicle control: saline. At termination, collect liver and spleen, weigh, and digest in nitric acid for iron measurement by atomic absorption spectroscopy or inductively coupled plasma mass spectrometry (ICP-MS). Stain liver and spleen sections with Perl's Prussian blue for iron visualization. Measure serum ferritin by ELISA. Hepcidin-treated mice should have lower tissue iron levels compared to iron-overload controls. For anemia of chronic disease models, inject mice with turpentine (50 uL intramuscularly) to induce inflammation. 24 hours later, treat with hepcidin (5 mg/kg, i.p.) for 7 days. Measure hemoglobin by automated hematology analyzer and serum iron. Hepcidin may exacerbate anemia. For bone studies, treat ovariectomized mice (osteoporosis model) with hepcidin (1-5 mg/kg, i.p., 3 times/week for 4 weeks). Assess bone mineral density by micro-CT and osteoclast activity by TRAP staining in tibial sections. All animal procedures require IACUC approval.
|
| ADME/Pharmacokinetics |
No specific pharmacokinetic (PK) data are available for Hepcidin-1 (mouse) TFA. As a 25-amino acid peptide (MW ~2.7 kDa), it is rapidly cleared from the circulation following intravenous or intraperitoneal administration. The plasma half-life (t1/2) in rodents is 5-15 minutes due to glomerular filtration (renal clearance) and proteolytic degradation. It does not cross the blood-brain barrier. It is not orally bioavailable. The peptide is primarily excreted in the urine as intact peptide and degraded fragments. The TFA salt does not affect PK. For a PK study, administer the peptide (2.5 mg/kg, i.v. or i.p.) to C57BL/6J mice (n=3-4 per time point). Collect blood at 0, 2, 5, 10, 15, 30, 60, 120, 240 minutes. Quantify hepcidin concentration by LC-MS/MS (using a stable isotope-labeled internal standard, e.g., Hepcidin-1-15N). PK parameters (AUC, Cmax, Tmax, t1/2, CL, Vd) are calculated using non-compartmental analysis. The short half-life necessitates multiple daily injections or continuous infusion for sustained pharmacodynamic effects.
|
| Toxicity/Toxicokinetics |
No specific toxicity data are available for Hepcidin-1 (mouse) TFA. As an endogenous peptide hormone, hepcidin-1 (mouse) is generally well-tolerated at physiological concentrations. In acute studies, doses up to 10 mg/kg i.p. in mice do not cause overt signs of toxicity (e.g., mortality, severe weight loss, behavioral abnormalities). At high doses (≥20 mg/kg), it may cause severe hypoferremia, leading to anemia (reduced hemoglobin and hematocrit) and possibly tissue iron deficiency, affecting erythropoiesis. Chronic elevation of hepcidin (e.g., in anemia of chronic disease) leads to iron-restricted anemia, but this is a pharmacological effect, not toxicity. No genotoxicity, carcinogenicity, or reproductive toxicity studies have been conducted. The TFA salt is present in low, stoichiometric amounts and is considered non-toxic. Standard laboratory safety precautions (gloves, lab coat, eye protection) should be used. Hepcidin-1 (mouse) TFA is for research use only and is not intended for human or therapeutic use.
|
| References | |
| Additional Infomation |
Hepcidin (encoded by the HAMP gene) is the master regulator of systemic iron homeostasis. It is a 25-amino acid cysteine-rich peptide that forms a hairpin structure with four disulfide bonds. Hepcidin is produced predominantly by hepatocytes in response to high iron levels, inflammation (via IL-6-STAT3 signaling), and infection. Its main target is ferroportin (FPN1), the only known cellular iron exporter. Hepcidin binds to ferroportin, leading to its internalization and degradation, thereby reducing iron export from enterocytes (dietary absorption), macrophages (iron recycling), and hepatocytes (iron storage). Hepcidin deficiency causes iron overload (e.g., hereditary hemochromatosis), while hepcidin excess leads to iron-restricted anemia (e.g., anemia of chronic disease, some forms of iron-refractory iron deficiency anemia). Hepcidin-1 (mouse) is the mouse ortholog of human hepcidin. It is widely used as a research tool to study iron metabolism disorders, anemia, inflammation, infection, and osteoporosis. The TFA salt is used to enhance peptide solubility and stability. As of 2026, hepcidin itself is not an approved drug, but hepcidin agonists and antagonists are in development for anemias and iron overload disorders. This product is for research use only and is not approved for human therapy.
|
| Molecular Formula |
C111H169N31O35S8.XC2HF3O2
|
|---|---|
| Molecular Weight |
2754.24 (free base)
|
| Related CAS # |
Hepcidin-1 (mouse);1676104-75-8
|
| Appearance |
Solid powder
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O :~100 mg/mL
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
