Description
Overview
| Product name: | Mouse ET-1(Endothelin1) ELISA Kit |
| Reactivity: | Mouse |
| Alternative Names: | ET1; PPET1; Preproendothelin-1; Big endothelin-1; EDN1 |
| Assay Type: | Sandwich |
| Sensitivity: | 2.43 pg/mL |
| Standard: | 500 pg/mL |
| Range: | 7.82-500 pg/mL |
| Sample Type: | serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
| Assay Length: | 3.5h |
| Research Area: | Endocrinology;Cardiovascular biology; |
| Test principle: | The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Endothelin1(ET-1). Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Endothelin1(ET-1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Endothelin1(ET-1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Endothelin1(ET-1) in the samples is then determined by comparing the OD of the samples to the standard curve. |
Standard curve
| Concentration (ng/mL) | OD | Corrected OD |
| 500.00 | 2.204 | 2.100 |
| 250.00 | 1.700 | 1.596 |
| 125.00 | 1.186 | 1.082 |
| 62.50 | 0.726 | 0.622 |
| 31.25 | 0.516 | 0.412 |
| 15.63 | 0.381 | 0.277 |
| 7.82 | 0.228 | 0.124 |
| 0.00 | 0.104 | 0.000 |
Precision
Intra-assay Precision (Precision within an assay)：CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays)：CV%<10%
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Precision
Intra-assay Precision (Precision within an assay)：CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays)：CV%<10%
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Recovery
Matrices listed below were spiked with certain level of recombinant ET-1 and the recovery rates were calculated by comparing the measured value to the expected amount of ET-1 in samples.
| Matrix | Recovery range | Average |
|---|---|---|
| serum(n=5) | 80-92% | 86% |
| EDTA plasma(n=5) | 85-97% | 91% |
| Heparin plasma(n=5) | 83-95% | 89% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of ET-1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
|---|---|---|---|---|
| serum(n=5) | 85-92% | 82-96% | 87-103% | 95-102% |
| EDTA plasma(n=5) | 78-96% | 89-102% | 85-97% | 79-97% |
| Heparin plasma(n=5) | 92-104% | 87-96% | 89-101% | 82-90% |
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