Description
The Intrinsic Mouse ERFETM ELISA is a test kit designed for quantification of mouse Erythroferrone in serum. This kit does not contain mercaptoethanol, azide- or mercury-based preservatives.
Detection of mouse erythroferrone is based on the double antibody sandwich ELISA method. The 96-well microplate assay plate is pre-coated with anti-mouse erythroferrone polyclonal antibody. Mouse serum samples, standards, or controls are mixed with buffer and incubated in the assay plate. Mouse erythroferrone present in the sample binds to the immobilized antibody on the assay plate. The plate is then washed to remove unbound mouse erythroferrone. A 2nd anti-mouse erythroferrone antibody conjugated with biotin is added and followed with HRP-SA conjugate which binds to biotinylated anti-mouse erythroferrone. Optical density (OD) is measured by the addition of TMB, the reaction produces a blue color and development is halted with the addition of the stop solution. The OD is read at 450nm on a microplate reader and a standard curve is prepared by plotting the log concentration of the standard curve versus its corresponding OD. The intensity of the color is proportional to the concentration of ERFE in the sample. Total assay run time is less than 3 hours.
Note:
Based on our studies, serum ERFE concentrations in healthy mice (C57BL/6) were essentially undetectable (below the limit of detection). In a mouse model of ineffective erythropoiesis (beta-thalassemia), serum ERFE concentrations ranged from 1-2 ng/ml.
ERF-200 Product Insert
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General Information
| Catalog number |
ERF-200 |
| Quantity |
96 Wells |
| Application |
ELISA, EIA, RUO |
| Method |
Sandwich |
| Antibody type |
Polyclonal |
| Target species |
Mouse |
| Analyte |
Erythroferrone |
| Assay time |
2h 45m |
| Detection method |
Colormetric |
| Sample type |
Serum |
| Sample volume required per well |
10 ul |
| Detection range |
0.0625 – 4.0 ng/ML |
| Sensitivity |
0.014 ng/mL |
| Regulatory Status |
Research use only ELISA |
| Storage requirements |
2-8 c |
| Shelf Life |
3 Months from Kit Assembly |
Typical data
| Erythroferrone Standard |
Concentration (ng/ML) |
Absorbance (450nm) |
| 1 |
4 |
2.196 |
| 2 |
2 |
1.321 |
| 3 |
1 |
0.727 |
| 4 |
0.5 |
0.410 |
| 5 |
0.25 |
0.243 |
| 6 |
0.125 |
0.146 |
| 7 |
0.063 |
0.100 |
| 8 |
0.001 |
0.056 |
Precision
|
Intra-assay Precision |
Inter-assay Precision |
| Sample |
1 |
2 |
3 |
1 |
2 |
3 |
| n |
16 |
16 |
16 |
16 |
16 |
16 |
| Mean(ng/ml) |
0.5 |
6.2 |
15.3 |
0.5 |
6.0 |
14.5 |
| Standard deviation |
0.03 |
0.26 |
0.56 |
0.03 |
0.35 |
0.63 |
| CV (%) |
5.6 |
4.2 |
3.7 |
7.2 |
5.7 |
4.3 |
Linearity
|
Serum |
| 1:2 |
Range (%) |
90-111 |
|
Average (%) |
99 |
| 1:4 |
Range (%) |
97-123 |
|
Average (%) |
106 |
| 1:8 |
Range (%) |
96-117 |
|
Average (%) |
109 |
Recovery
| Sample Type |
Range (%) |
Average Recovery (%) |
| Serum |
89-109 |
101 |
