8-hydroxy-2-deoxyguanosine (8-OHdG) ELISA


SKU: HDG39-K01 Categories: , ,
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8-hydroxy-2′-deoxyguanosine (8-OHdG) ELISA Assay kit: 
For Research Use Only
Size:  1 x 96 wells
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

Intended Use
The Eagle Biosciences 8-hydroxy-2′-deoxyguanosine (8-OHdG) 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. 

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.

  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. Incubate at room temperature for 15 minutes in the dark.
  11. Add 100µL of the Reaction Terminating Solution. Shake the plate from side to side and mix fully.
  12. Measure the absorbance at 450 nm using microtiter plate reader.

Typical Standard Curve:



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