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Human Hepcidin Quantikine ELISA Kit

R&D Systems | Catalog # DHP250

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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)
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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

Species

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.

Sample TypeMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL
Serum (n=40)18,36579.0-49,40014,677
EDTA plasma (n=40)19,39982.4-56,70015,506
Heparin plasma (n=40)17,46783.3-49,20114,191

Sample TypeMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Urine (n=12)79,0042104-377,482111,333
Note: Normalization to urine creatinine (UCr) levels is recommended when assaying urine samples.

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.

Human Hepcidin ELISA Linearity

Scientific Data Images for Human Hepcidin Quantikine ELISA Kit

Human Hepcidin ELISA Standard Curve

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

The product encoded by this gene is involved in the maintenance of iron homeostasis, and it is necessary for the regulation of iron storage in macrophages, and for intestinal iron absorption. The preproprotein is post-translationally cleaved into mature peptides of 20, 22 and 25 amino acids, and these active peptides are rich in cysteines, which form intramolecular bonds that stabilize their beta-sheet structures. These peptides exhibit antimicrobial activity. Mutations in this gene cause hemochromatosis type 2B, also known as juvenile hemochromatosis, a disease caused by severe iron overload that results in cardiomyopathy, cirrhosis, and endocrine failure.

Long Name

Hepcidin Antimicrobial Peptide

Alternate Names

HAMP, HEPC, HFE2B, LEAP1, PLTR

Entrez Gene IDs

57817 (Human); 84506 (Mouse); 84604 (Rat)

Gene Symbol

HAMP

Additional Hepcidin Products

Product Documents for Human Hepcidin Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Hepcidin Quantikine ELISA Kit

For research use only

⚠ 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.

Related Research Areas

Red Blood Cells (RBCs)

Citations for Human Hepcidin Quantikine ELISA Kit

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    Sample Tested: human floxp embryonal stem cell culture medium
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    Human Hepcidin Quantikine ELISA Kit DHP250

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