Cardiovascular and Oxidative Stress Assay Kit

8-hydroxy-2-deoxyguanosine ELISA Assay Kit

$1,575.00

The 8-hydroxy-2-deoxyguanosine ELISA Assay Kit 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-hydroxy-2′-deoxyguanosine (8-OHdG) ELISA assay kit is for research use only and not to be used in diagnostic procedures. 

SKU: HDG39-K01 Categories: , ,

8-hydroxy-2-deoxyguanosine ELISA Assay Kit

The 8-hydroxy-2-deoxyguanosine ELISA Assay Kit is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.5 ng/mL
Dynamic Range: 0.5 – 200 ng/mL
Incubation Time: 3.5 hours
Sample Type: Serum, Urine, Biological Fluids
Sample Size: 50 µL
Alternative Names: 8-OHdG

Assay Principle

Bring all reagents to room temperature before beginning 8-hydroxy-2′-deoxy-guanosine (8-OHdG) 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 anti-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 HRP-conjugated secondary antibody is removed by washing.
Addition of the substrate solution results in the development of color in proportion to the amount of anti 8-OHdG antibody bound to the plate.
The reaction is terminated by phosphoric acid, and absorbance at 450 nm is measured.

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Additional Information

Assay Principle

Bring all reagents to room temperature before beginning 8-hydroxy-2′-deoxy-guanosine (8-OHdG) 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.

Assay Procedure

  1. Reconstitute the Primary Antibody with the Primary Antibody Solution.
  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 37°C for 1 hour.
  4. Measured values may be very much affected with the incubation temperatures, particularly during primary antibody reaction period.  It is recommended to use water bath rather than dry incubators for the incubation.
  5. Mix 1 volume of Washing Solution (5x) with 4 volumes of distilled water.
  6. Pour off contents of wells 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. The use of washing machines or aspirators is not recommended.
  7. Reconstitute the Secondary Antibody with the Secondary Antibody Solution.
  8. Add 100µL of reconstituted secondary antibody per well. Shake the plate from side to side and mix fully. Cover the plate with an adhesive strip. Incubate 37°C for 1 hour.
  9. At the end of the incubation period, repeat washing as in step 5.
  10. Prepare substrate solution. Add 1 volume of Chromatic Solution to 100 volumes of Diluting Solution just before use. Add 100µL of substrate solution per well. Shake the plate from side to side and mix fully.
  11. Incubate at room temperature for 15 minutes in the dark.
  12. Add 100µL of the Reaction Terminating Solution. Shake the plate from side to side and mix fully.
  13. Measure the absorbance at 450 nm using microtiter plate reader.

Typical Standard Curve

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