Cardiovascular and Oxidative Stress Assay Kit

8-OHdG Ultrasensitive ELISA Assay

$1,575.00

The 8-OHdG Ultrasensitive ELISA Assay is intended for the quantitative determination of adduct 8-hydroxy-2’-deoxy-guanosine (8-OHdG) in urine, serum or biological samples by enzyme linked immunoassay (ELISA). The 8-OHdG Ultrasensitive ELISA Assay kit is for research use only and not to be used in diagnostic procedures.

SKU: HDU39-K01 Categories: , ,

8-OHdG Ultrasensitive ELISA Assay

The 8-OHdG Ultrasensitive ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.125 ng/mL
Dynamic Range: 0.125 – 10 ng/mL
Incubation Time: Overnight
Sample Type: Serum, Urine, Biological Fluids
Sample Size: 50 µL
Alternative Names: 8-hydroxy-2’-deoxy-guanosine

Product manufactured in the USA

Assay Principle

Bring all reagents of the  to room temperature before beginning 8-hydroxy-2′-deoxy-guanosine (8-OHdG) Ultrasensitive ELISA assay kit.  Determine the number of microwells needed for the assay (each sample, standard, and control should be assayed in duplicate).  Bring all reagents and samples to room temperature (20-­25ºC) before use.
The 8-OHdG monoclonal antibody and the sample or standard are added to the microtiter plate which has been precoated with 8-OHdG. The 8-OHdG monoclonal antibody reacts competitively with the 8-OHdG bound on the plate and the 8-OHdG in samples solution. Therefore higher concentrations of 8-OHdG in the sample solution lead to a reduced binding of the antibody to the 8-OHdG on the plate.
The antibodies which are bound to the 8-OHdG in the sample are washed away from the antibodies that have bound to the 8-OHdG coated on the plate.
An enzyme-labeled secondary antibody, which is added to the plate, binds to the monoclonal antibody which is bound to the 8-OHdG coated on the plate.
Unbound enzyme-labeled secondary antibody is removed by a wash step.
Addition of a chromatic substrate results in the development of color in proportion to the amount of antibody bound to the plate.
The color reaction is terminated and the absorbance is measured.

Related Products

8-OHdG ELISA Assay Kit
8-hydroxy-2-deoxyguanosine ELISA Assay Kit
Hexanoyl-Lys adduct ELISA Assay

Additional Information

Assay Procedure

  1. Reconstitute the primary antibody with the primary antibody solution. Allow dissolving completely.
  2. Add 50µl of sample or standard per well.
  3. Add 50µl of reconstituted primary antibody per well. Shake the plate from side to side and mix fully. Cover plate with adhesive strip, making sure it is sealed tightly. Incubate at 4C for overnight.
  4. Pour off contents of plate into sink. Pipette 250µl washing solution into each well. After washing thoroughly by shaking the plate from side to side, dispose of washing solution. Invert plate and blot against clean paper towel to remove any remaining washing buffer. Repeat wash two times more.
  5. Reconstitute the secondary antibody with the secondary antibody solution. Allow dissolving completely.
  6. Add 100µl of constituted secondary antibody per well. Shake the plate from side to side and mix fully.
  7. Cover the plate with an adhesive strip. Incubate room temperature for 1 hour.
  8. At the end of the incubation period, repeat wash as in step 4.
  9. Reconstitute the chromatic solution (enzyme substrate solution) with 100 times volume of the diluting solution. Add 100µl of the reconstituted enzyme substrate per well. Shake the plate from side to side and mix fully. Incubate at room temperature for 15 minutes. This incubation should be done in the dark, i.e. shield the plate with aluminum foil.
  10. Add 100µl of the reaction terminating solution. Shake the plate from side to side and mix fully.
    After terminating the reaction, read the absorbance at 450 nm.
  11. Use a standard curve to determine the amount of 8-OHdG present in test samples. Generate the standard curve by plotting absorbance vs. log (concentration of standards). Then use the absorbance values obtained for the test samples to determine the concentrations.

Typical Standard Curve

8-OHdG Ultrasensitive ELISA Assay

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