Human Hepcidin (Hepc) ELISA Kit

The Human Hepcidin ELISA Kit is a sensitive and specific assay designed for the quantitative detection of Hepcidin levels in human serum, plasma, urine, culture media, and other biological fluids. Hepcidin is a key regulator of iron metabolism, and this kit is essential for research in iron homeostasis, anemia, and related disorders.

Sample Preparation

1. Serum Preparation

1. Collect whole blood and allow it to clot at room temperature for 10–20 minutes.
2. Centrifuge at 2,000–3,000 rpm for 20 minutes to separate the serum.
3. Re-centrifuge if precipitates appear during storage.

2. Plasma Preparation

1. Collect whole blood into tubes containing anticoagulants (e.g., EDTA or citrate).
2. Incubate at room temperature for 10–20 minutes.
3. Centrifuge at 2,000–3,000 rpm for 20 minutes to obtain plasma.
4. Re-centrifuge if necessary.

3. Urine Samples

1. Collect urine in aseptic tubes.
2. Centrifuge at 2,000–3,000 rpm for 20 minutes and carefully collect the supernatant.
3. Re-centrifuge if precipitates form during storage.

4. Cell Samples

1. For cell secretions, collect culture supernatants in aseptic tubes and centrifuge at 2,000–3,000 rpm for 20 minutes.
2. For intracellular components:
   • Dilute cells in PBS (pH 7.2–7.4) at 1×10⁷/mL.
   • Disrupt cells via repeated freeze-thaw cycles.
   • Centrifuge at 2,000–3,000 rpm for 20 minutes and collect the supernatant.

5. Tissue Samples

1. Cut and weigh tissue samples, then freeze in liquid nitrogen and store at -80°C.
2. Homogenize samples in PBS (pH 7.4) at 4°C.
3. Centrifuge at 2,000–3,000 rpm for 20 minutes and collect the supernatant.

Important Notes

1. Perform ELISA assays as soon as possible after sample collection.
2. Store samples at -20°C if immediate testing is not feasible.
3. Avoid repeated freeze-thaw cycles to preserve sample integrity.
4. Do not use samples containing sodium azide (NaN₃) as it inhibits HRP activity.

Applications

• Iron Homeostasis Research:
  • Study Hepcidin as a key regulator of iron absorption and distribution.
• Anemia and Iron Disorders:
  • Investigate Hepcidin levels in conditions such as anemia of chronic disease, iron deficiency anemia, and hemochromatosis.
• Metabolic Studies:
  • Explore the relationship between Hepcidin and metabolic disorders.
• Inflammation Research:
  • Study the role of Hepcidin as an acute-phase reactant in inflammatory conditions.
• Therapeutic Development:
  • Evaluate drugs targeting iron metabolism and Hepcidin regulation.

Advantages

1. High Sensitivity and Specificity:
   • Detects low levels of Hepcidin with minimal cross-reactivity or interference.
2. Wide Sample Compatibility:
   • Validated for serum, plasma, urine, cell culture supernatants, and tissue homogenates.
3. Ease of Use:
   • Includes pre-coated plates and ready-to-use reagents.
4. Reproducibility:
   • Provides consistent and reliable results across multiple experiments.

Storage and Stability

• Store all kit components at 2–8°C.
• Avoid repeated freeze-thaw cycles to ensure assay reliability.

The Human Hepcidin ELISA Kit is a vital tool for researchers studying iron metabolism, anemia, and related conditions. Its precision, user-friendly design, and broad compatibility make it indispensable for various clinical and experimental applications.