Description
The Intrinsic Rat Hepcidin™ ELISA is a test kit designed for quantification of rat Hepcidin in serum.
This kit does not contain mercaptoethanol, azide- or mercury-based preservatives.
This kit is a solid-phase enzyme-linked immunosorbent assay (ELISA), based on the principles of competitive binding. Serum samples, standards, or controls are mixed with buffer containing rat hepcidin biotin conjugate. This mixture is incubated in an anti-rat hepcidin antibody coated ELISA microwell plate. The more rat hepcidin in the sample, the less rat hepcdin biotin conjugate will bind to the antibody coated well due to “competition” for antibody binding sites between native rat hepcidin and synthetic rat hepcidin biotin conjugate. The plate is then washed to remove unbound rat hepcidin biotin conjugate. The rat hepcidin biotin conjugate bound to the antibody is detected with streptavidin conjugated horseradish peroxidase (HRP). The binding of the rat hepcidin biotin conjugate is quantified by the addition of TMB, a chromogenic substrate. The reaction produces a blue color and development is stopped with the addition of a stop solution and the absorbance is read at 450 nm. A standard curve is produced by plotting the concentration of the standard versus the absorbance. The intensity of the color is inversely proportional to the concentration of rat hepcidin in the sample. The total assay run time is less 3 hours.
RHC-007 Product Insert
Researcher’s Guide to Hepcidin Testing
Questions? Please email us: [email protected]
General Information
| Catalog number |
RHC-007 |
| Quantity |
96 Wells |
| Application |
ELISA, EIA, RUO |
| Method |
Competitive |
| Antibody type |
Polyclonal |
| Target species |
Rat |
| Analyte |
Hepcidin |
| Assay time |
2h 45m |
| Detection method |
Colorimetric |
| Sample type |
Serum |
| Sample volume required per well |
6ul |
| Detection range |
1.37-330 ng/ml |
| Sensitivity |
LLOD: 0.86ng/ml LLOQ: 2.48ng/ml |
| Regulatory Status |
Research Use Only ELISA |
| Storage requirements |
2-8 °c |
| Shelf Life |
4 Months from Kit Assembly |
Typical data
| Hepcidin Standard |
Concentration (ng/ML) |
Absorbance (450nm) |
| 1 |
1000 |
0.162 |
| 2 |
333.3 |
0.275 |
| 3 |
111.1 |
0.531 |
| 4 |
37.0 |
1.185 |
| 5 |
12.3 |
1.918 |
| 6 |
4.1 |
2.424 |
| 7 |
1.4 |
2.741 |
| 8 |
0 |
2.836 |
Precision
|
Intra-assay Precision |
Inter-assay Precision |
| Sample |
1 |
2 |
3 |
1 |
2 |
3 |
| n |
24 |
24 |
24 |
24 |
24 |
24 |
| Mean(ng/ml) |
84.3 |
169.7 |
594.4 |
84.3 |
169.7 |
594.4 |
| Standard deviation |
11.6 |
18.3 |
43.6 |
13.6 |
21.6 |
50.3 |
| CV (%) |
14 |
11 |
7 |
16 |
13 |
8 |
Linearity
|
Serum |
| 1:2 |
Range (%) |
96-102% |
|
Average (%) |
99% |
| 1:4 |
Range (%) |
103-107% |
|
Average (%) |
105% |
Recovery
| Sample Type |
Range (%) |
Average Recovery (%) |
| Serum |
93-107 |
102 |
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