Lablisa® Human 2,3-DPG(2,3-Diphosphoglycerate) ELISA Kit
Product Information
- Description
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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 Human 2,3-DPG. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human 2,3-DPG. 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 Human 2,3-DPG, 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 Human 2,3-DPG in the samples is then determined by comparing the OD of the samples to the standard curve.
| Product name: | Human 2,3-DPG(2,3-Diphosphoglycerate) ELISA Kit |
| Reactivity: | Human |
| Alternative Names: | 2,3DPG |
| Assay Type: | Sandwich |
| Sensitivity: | 0.14 μmol/mL |
| Standard: | 20 μmol/mL |
| Detection Range: | 0.32-20 μmol/mL |
| Sample Type: | Serum, plasma, tissue homogenates, cell lysates and other biological fluids |
| Assay Length: | 3.5h |
| Research Area: | Metabolic pathway |
Standard curve
| Concentration (μmol/mL) | OD | Corrected OD |
|---|---|---|
| 20.00 | 2.154 | 2.059 |
| 10.00 | 1.587 | 1.492 |
| 5.00 | 1.184 | 1.089 |
| 2.50 | 0.866 | 0.771 |
| 1.25 | 0.519 | 0.424 |
| 0.63 | 0.336 | 0.241 |
| 0.32 | 0.190 | 0.095 |
| 0.00 | 0.095 | 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.
Recovery
Matrices listed below were spiked with certain level of recombinant 2,3-DPG and the recovery rates were calculated by comparing the measured value to the expected amount of 2,3-DPG in samples.
| Matrix | Recovery range | Average |
|---|---|---|
| serum(n=5) | 90-105% | 97% |
| EDTA plasma(n=5) | 80-92% | 86% |
| Heparin plasma(n=5) | 88-102% | 95% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 2,3-DPG 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) | 85-97% | 90-99% | 82-101% | 93-105% |
| EDTA plasma(n=5) | 85-97% | 87-96% | 82-95% | 92-105% |
| Heparin plasma(n=5) | 87-96% | 92-105% | 89-102% | 92-101% |
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