INFORMATION
| Full Name | 8-Hydroxydeoxyguanosine |
|---|---|
| Synonym | 8OHdG, 8-Oxo-DG, 7, 8-Dihydro-8-Oxo-2'-Deoxyguanosine |
| Assay Type | Competitive |
| Reactivity | Universal |
| Range | 1.56-100ng/mL |
| Sensitivity | 0.94ng/mL |
| Sample Type | Serum, plasma and other biological fluids |
| Sample Volume | 50μL |
| Detection Wavelength | OD450 |
| Reaction Duration | 1.5h |
| Transportation Temperature | 2-8°C |
| Specificity | Specifically binds with 8-OHdG, no obvious cross reaction with other analogues. |
| Microplate | 96-wells plate breakable into 12 x 8 wells strip |
ELISA KIT COMPONENTS
Upon receipt, unpack promptly and store as recommended in the instructions.
| Components | Specifications | Storage and Notes |
|---|---|---|
| Micro Plate | 96T: 8 wells×12 strips 48T: 8 wells×6 strips | Unopened: -20°C, 12 months Unused: Put it back in the aluminum foil bag and seal it, store it at -20°C. |
| Reference Standard | 96T: 2 vials 48T: 1 vial | Unopened: -20°C, 12 months Please use freshly dissolved standards for each experiment. Discard any unused standards after dissolution. |
| Biotinylated Detection Ab Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C, 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| HRP Conjugate Concentrate (100×) | 96T: 120μL×1 vial 48T: 60μL×1 vial | Unopened: -20°C(Protect from light), 12 months Unused: Please seal the concentrate and store it at -20°C, and discard the working solution. |
| Biotinylated Detection Ab Diluent | 14mL×1 | 2-8℃, 12 months |
| HRP Conjugate Diluent | 14mL×1 | 2-8℃, 12 months |
| Reference Standard & Sample Diluent | 20mL×1 | 2-8℃, 12 months |
| Washing Buffer Concentrate (25×) | 30mL×1 | 2-8℃, 12 months |
| Substrate Reagent(TMB) | 10mL×1 | 2-8°C(Protect from light),12 months |
| Stop Solution | 7mL×1 | 2-8°C/Room temperature |
REQUIRED INSTRUMENTS AND REAGENTS
- Microplate reader (450nm wavelength filter).
- Incubator capable of maintaining 37℃.
- 1.5mL EP tube, absorbent paper.
- Pipette and disposable tip: 0.5-10μL, 2-20μL, 20-200μL, 200-1000μL.
- Double distilled water or deionized water.
ASSAY PROCEDURE SUMMARY
STANDARD CURVE
| STD.(ng/mL) | OD-1 | OD-2 | Average |
|---|---|---|---|
| 0 | 2.174 | 2.176 | 2.175 |
| 1.57 | 1.846 | 1.852 | 1.849 |
| 3.13 | 1.591 | 1.567 | 1.579 |
| 6.25 | 1.226 | 1.21 | 1.218 |
| 12.5 | 0.841 | 0.863 | 0.852 |
| 25 | 0.583 | 0.561 | 0.572 |
| 50 | 0.398 | 0.402 | 0.4 |
| 100 | 0.306 | 0.31 | 0.308 |
RECOVERY
The recovery of 8-OHdG spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
LINEARITY
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of 8-OHdG in various matrices were diluted with the Reference Standard & Sample Diluent to produce samples with values within the dynamic range of the assay.
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.
| Item | Intra-assay Precision | Inter-assay Precision | ||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 5.09 | 12.87 | 48.53 | 5.08 | 12.31 | 50.74 |
| Standard deviation | 0.28 | 0.61 | 2.45 | 0.34 | 0.54 | 2.07 |
| CV(%) | 5.51 | 4.71 | 5.04 | 6.61 | 4.4 | 4.08 |
STABILITY
Perform stability testing of the kit at 37°C and 2-8°C and obtain relevant data.
| ELISA kit(n=5) | 37°C for 1 month | 2-8°C for 6 months |
|---|---|---|
| Average(%) | 80 | 95-100 |
