8-OHdG ELISA Assay Kit

$490.00$2,090.00

The 8-OHdG ELISA Assay Kit is a competitive assay that can be used for the quantification of 8-OHdG in urine, cell culture, plasma, and other sample matrices.

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SKU: SKT-120 Categories: , ,

8-OHdG ELISA Assay Kit

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

Sensitivity: 0.59 ng/mL
Dynamic Range: 0.94 – 60 ng/mL
Incubation Time: 1 hour
Sample Type: Cell lysates, Plasma, Sample matrices, Urine
Sample Size: 50 μL

Product manufactured in Canada by StressMarq.


Assay Background

8-hydroxy-2-deoxy Guanosine (8-OH-dG) is produced by the oxidative damage of DNA by reactive oxygen and nitrogen species and serves as an established marker of oxidative stress. Hydroxylation of guanosine occurs in response to both normal metabolic processes and a variety of environmental factors (i.e., anything that increases reactive oxygen and nitrogen species). Increased levels of 8-OH-dG are associated with the aging process as well as with a number of pathological conditions including cancer, diabetes, and hypertension. In complex samples such as plasma, cell lysates, and tissues, 8-OH-dG can exist as either the free nucleoside or incorporated in DNA. Once the blood enters the kidney, free 8-OH-dG is readily filtered into the urine, while larger DNA fragments remain in the bloodstream. Because of the complexity of plasma samples, urine is a more suitable matrix for the measurement of free 8-OH-dG than plasma. Urinary levels of 8-OH-dG range between 2.7-13 ng/mg creatine, while plasma levels of free 8-OH-dG have been reported to be between 4-21 pg/ml as determined by LC-MS.


Storage and Stability
This kit will perform as specified if the components are stored as directed and used before the expiration date indicated on the outside of the box. All reagents are stable as supplied at 4°C, except the standard, which should be stored at -20°C. For optimum storage, the 8-OHdG Standard should be aliquoted into smaller portions and then stored appropriately. Avoid repeated freeze/thaw cycles (10 µL of Standard can prepare a triplicate standard curve.


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Additional Information

Assay Procedure


  1. Prepare standard and samples in the Sample and Standard Diluent.
  2. Add 50 µL of prepared standards and samples in triplicate to appropriate wells.
  3. Add 50 µL of the diluted antibody preparation to the appropriate wells.
  4. Cover plate with Plate Cover and incubate at room temperature (20-25°C) for 1 hour.
  5. Wash plate 4 times with 1X Wash Buffer.
  6. Add 100 µL of TMB Substrate to each well.
  7. Cover plate and develop the plate in the dark at room temperature for 30 minutes.
  8. Add 100 µL of Stop Solution to each well.
  9. Measure absorbance on a plate reader at 450 nm.
  10. Plot the standard curve and calculate sample concentrations.

Typical Standard Curve


8-OHdG ELISA Assay Kit

Manual

Product Manual