Description
8-OHdG (8-Hydroxydeoxyguanosine) ELISA Kit
This ELISA kit uses Competitive-ELISA as the method. The microtiter plate provided in this kit has been pre-coated with 8-OHdG. During the reaction, 8-OHdG in the sample or standard competes with a fixed amount of 8-OHdG on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 8-OHdG. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of 8-OHdG in the samples is then determined by comparing the OD of the samples to the standard curve.
Assay type | Competitive-ELISA |
Format | 96T |
Assay time | 2.5h |
Reactivity | Universal |
Detection Method | Colormetric |
Detection Range | 1.56-100 ng/mL |
Sensitivity | 0.94 ng/mL |
Sample Volume | 50uL |
Sample Type | Serum, plasma and other biological fluids |
Specificity
This kit recognizes 8-OHdG in samples. No significant cross-reactivity or interference between 8-OHdG and analogues was observed.
Typical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Concentration(ng/mL) | O.D | Average |
---|---|---|
100 | 0.398 0.454 | 0.426 |
50 | 0.51 0.562 | 0.536 |
25 | 0.734 0.724 | 0.729 |
12.5 | 1.014 1.034 | 1.024 |
6.25 | 1.387 1.385 | 1.386 |
3.13 | 1.746 1.718 | 1.732 |
1.56 | 1.993 1.993 | 1.993 |
0 | 2.325 2.337 | 2.331 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level 8-OHdG were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level 8-OHdG were tested on 3 different plates, 20 replicates in each plate.
Intra-assay Precision | Inter-assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean (ng/mL) | 4.77 | 8.88 | 45.84 | 4.81 | 9.56 | 43.89 |
Standard deviation | 0.24 | 0.49 | 2.27 | 0.34 | 0.49 | 1.94 |
C V (%) | 5.03 | 5.52 | 4.95 | 7.07 | 5.13 | 4.43 |
Recovery
The recovery of 8-OHdG spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type | Range (%) | Average Recovery (%) |
---|---|---|
Serum (n=5) | 90-105 | 97 |
EDTA plasma (n=5) | 92-105 | 98 |
Cell culture media (n=5) | 84-95 | 90 |
Linearity
Samples were spiked with high concentrations of 8-OHdG and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
---|---|---|---|---|
1:2 | Range (%) | 97-111 | 96-109 | 96-111 |
Average (%) | 103 | 101 | 103 | |
1:4 | Range (%) | 94-105 | 89-105 | 101-113 |
Average (%) | 99 | 96 | 106 | |
1:8 | Range (%) | 93-108 | 97-114 | 98-110 |
Average (%) | 99 | 104 | 104 | |
1:16 | Range (%) | 96-111 | 93-108 | 93-107 |
Average (%) | 102 | 99 | 99 |
Kit Components & Storage
An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
Item | Specifications | Storage |
---|---|---|
Micro ELISA Plate(Dismountable) | 8 wells X12 strips | -20'C, 6 months |
Reference Standard | 2 vials | |
Concentrated Biotinylated Detection Ab (100X) | 1 vial, 120 uL | |
Concentrated HRP Conjugate (100X) | 1 vial, 120 uL | -20'C(shading light), 6 months |
Reference Standard & Sample Diluent | 1 vial, 20 mL | 4'C, 6 months |
Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
HRP Conjugate Diluent | 1 vial, 14 mL | |
Concentrated Wash Buffer (25X) | 1 vial, 30 mL | |
Substrate Reagent | 1 vial, 10 mL | 4'C(shading light) |
Stop Solution | 1 vial, 10 mL | 4'C |
Plate Sealer | 5 pieces | |
Product Description | 1 copy | |
Certificate of Analysis | 1 copy |
Additional Information
Product type: |
ELISA |
Reactivity: |
Universal |
ELISA Type: |
Competitive |