Human Hepcidin ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # DHP250
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (25 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Urine (10 µL)
Sensitivity
3.81 pg/mL
Assay Range
15.6-1000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Product Summary for Human Hepcidin Quantikine ELISA Kit
The Quantikine Human Hepcidin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Hepcidin in cell culture supernates, serum, plasma, and urine. It contains synthetic Hepcidin and antibodies raised against synthetic Hepcidin. This immunoassay has been shown to accurately quantitate synthetic and naturally occurring human Hepcidin.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Natural human Hepcidin.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma/Urine - Samples from apparently healthy volunteers were evaluated for the
presence of human Hepcidin in this assay. No medical histories were available for the donors
used in this study.
Note: Normalization to urine creatinine (UCr) levels is recommended when assaying urine samples.
| Sample Type | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL |
| Serum (n=40) | 18,365 | 79.0-49,400 | 14,677 |
| EDTA plasma (n=40) | 19,399 | 82.4-56,700 | 15,506 |
| Heparin plasma (n=40) | 17,467 | 83.3-49,201 | 14,191 |
| Sample Type | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Urine (n=12) | 79,004 | 2104-377,482 | 111,333 |
Cell Culture Supernates:
HepG2 human hepatocellular carcinoma cells were cultured in MEM supplemented with
10% fetal bovine serum, 1 mM sodium pyruvate, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. Cells were unstimulated or stimulated with 10 ng/mL of
recombinant human IL-6, 10 ng/mL of recombinant human IL-1 beta, and 10 ng/mL of
recombinant human TNF-alpha for 3 days. Aliquots of the cell culture supernates were removed,
assayed for human Hepcidin, and measured 4455 pg/mL and 20,000 pg/mL, respectively.
HepG2 human hepatocellular carcinoma cells were cultured in MEM supplemented with
10% fetal bovine serum, 1 mM sodium pyruvate, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate under normoxic conditions or in a hypoxia workstation at
1% O2 for 3 days. Aliquots of the cell culture supernates were removed, assayed for human
Hepcidin, and measured 1804 pg/mL and 3540 pg/mL, respectively.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 175 | 350 | 726 | 164 | 328 | 647 |
| Standard Deviation | 7.51 | 10.8 | 23.5 | 18.0 | 26.1 | 39.8 |
| CV% | 4.3 | 3.1 | 3.2 | 11.0 | 8.0 | 6.2 |
Recovery for Human Hepcidin Quantikine ELISA Kit
The recovery of human Hepcidin spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 111 | 99-118 |
| EDTA Plasma (n=4) | 103 | 95-115 |
| Heparin Plasma (n=4) | 104 | 93-117 |
| Serum (n=4) | 106 | 96-118 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of human Hepcidin were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Scientific Data Images for Human Hepcidin Quantikine ELISA Kit
Human Hepcidin ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Hepcidin
Long Name
Hepcidin Antimicrobial Peptide
Alternate Names
HAMP, HEPC, HFE2B, LEAP1, PLTR
Gene Symbol
HAMP
Additional Hepcidin Products
Product Documents for Human Hepcidin Quantikine ELISA Kit
Product Specific Notices for Human Hepcidin Quantikine ELISA Kit
For research use only
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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.