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

• Dilution of Standards

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