Mouse 8-Hydroxy-desoxyguanosine;8-OHdG ELISA Kit
- Our Mouse 8-Hydroxy-desoxyguanosine (8-OHdG) ELISA kit is to assay 8-OHdG levels in Mouse serum, plasma, culture media or any biological fluid.
Principle
This ELISA kit uses Sandwich-ELISA as the method. The Microelisa stripplate provided in this kit has been pre-coated with an antibody specific to 8-OHdG. Standards or samples are added to the appropriate Microelisa stripplate wells and combined to the specific antibody. Then a Horseradish Peroxidase (HRP)- conjugated antibody specific for 8-OHdG is added to each Microelisa stripplate well and incubated. Free components are washed away. The TMB substrate solution is added to each well. Only those wells that contain 8-OHdG and HRP conjugated 8-OHdG antibody will appear blue in color and then turn yellow after the addition of the stop solution. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm. The OD value is proportional to the concentration of 8-OHdG. You can calculate the concentration of 8-OHdG in the samples by comparing the OD of the samples to the standard curve.
Materials provided with the kit
| | Materials provided with the kit | 48 determinations | 96 determinations | Storage |
| 1 | User manual | 1 | 1 | R.T. |
| 2 | Closure plate membrane | 2 | 2 | R.T. |
| 3 | Sealed bags | 1 | 1 | R.T. |
| 4 | Microelisa stripplate | 1 | 1 | 2-8℃ |
| 5 | Standard 1350 pg/ml | 0.5ml×1 bottle | 0.5ml×1 bottle | 2-8℃ |
| 6 | Standard diluent | 1.5ml×1 bottle | 1.5ml×1 bottle | 2-8℃ |
| 7 | HRP-Conjugate reagent | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 8 | Sample diluent | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 9 | Chromogen Solution A | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 10 | Chromogen Solution B | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 11 | Stop Solution | 3ml×1 bottle | 6ml×1 bottle | 2-8℃ |
| 12 | Wash solution | 20ml (20X)×1bottle | 20ml (30X)×1bottle | 2-8℃ |
Ten wells are set for standards in a Microelisa stripplate. In Well 1 and Well 2, 100μl Standard solution and 50μl Standard Dilution buffer are added and mixed well. In Well 3 and Well 4, 100μl solution from Well 1 and Well 2 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. 50μl solution is discarded from Well 3 and Well 4. In Well 5 and Well 6, 50μl solution from Well 3 and Well 4 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. In Well 7 and Well 8, 50μl solution from Well 5 and Well 6 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. In Well 9 and Well 10, 50μl solution from Well 7 and Well 8 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. 50μl solution is discarded from Well 9 and Well 10. After dilution, the total volume in all the wells are 50μl and the concentrations are 900 pg/ml,600 pg/ml,300 pg/ml, 150 pg/ml and 75pg/ml, respectively.
Calculation of Results
Known concentrations of Mouse 8-OHdG Standard and its corresponding reading OD is plotted on the log scale (x-axis) and the log scale (y-axis) respectively. The concentration of Mouse 8-OHdG in sample is determined by plotting the sample’s O.D. on the Y-axis. The original concentration is calculated by multiplying the dilution factor.
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mouse 8-OHdG were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mouse 8-OHdG were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Assay range
15 pg/ml- 1000 pg/ml
Sensitivity:
4.5pg/ml
Product Tags:
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Sandwich 8-OHdG ELISA Kit Mouse 8-Hydroxydeoxyguanosine Test Kit Images
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