8-Isoprostane ELISA Assay Kit

$330.00$2,620.00

The Eagle Biosciences 8-Isoprostane ELISA Assay kit is intended for the quantitative determination of 8-isoprostane in biological samples by enzyme linked immunoassay (ELISA). 8-Isoprostane ELISA Assay kit is for research use only and not to be used in diagnostic procedures.

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SKU: 8IS39-K01 0 Categories: , ,

8-Isoprostane ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 10 pg/ml
Dynamic Range: 10 -5000 pg/ml
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma, Tissue, Cell Culture
Sample Size: 100 µl

Product manufactured in the USA

Additional Information

Assay Background


The isoprostanes are a family of eicosanoids of non-enzymatic origin produced by the random oxidation of tissues phospholipids by oxygen radicals.  A recent NIH-sponsored study on Biomarkers of Oxidative Stress has indicated that 8-isoprostane is the best index of oxidative injury in a well-accepted oxidant stress rat model (1,2).  In addition, plasma 8-isoprostane levels were found to be elevated in elderly subjects with severe hypertension (3) and in urine from subjects with high fat diet-induced liver steatosis (4).

This 8-Isoprostane ELISA Assay kit can be used for the determination of 8-isoprostane in diluted urine, serum, plasma, cells, and tissues following proper isolation and purification of the eicosanoid from the isoprostane-containing sample.  Instructions are provided as to the proper isolation and purification in the following pages.

Assay Principle


  1. Load 200 microliters of Sample Dilution Buffer into the blank (BL) wells and 100 microliters of Sample Dilution Buffer into the maximum binding (BO) wells.
  2. Load 100 microliters of each of the standards into the appropriate wells.
  3. Load 100 microliters of the diluted 8-isoprostane-HRP conjugate in the BO wells, the standard wells, and the sample wells.  Do NOT add HRP conjugate into the BL wells.
  4. Incubate the plate at room temperature for two hours.
  5. Wash the plate three times with 400 microliters of the diluted Wash Buffer per well.
  6. After the last of the three wash cycles pat the plate dry onto some paper toweling.
  7. Add 200 microliters of the TMB substrate to all of the wells (including BL wells)./li>
  8. Incubate the plate at room temperature for 15-30 minutes.
  9. Add 50 micoliters of 2 N sulfuric acid to all of the wells.
  10. Read the plate at 450 nm.

Typical Standard Curve


Manual

Product Manual


Publications

Citations


Bitzer, Zachary T, et al. “Effect of Flavoring Chemicals on Free Radical Formation in Electronic Cigarette Aerosols.” Free Radical Biology and Medicine, vol. 112, 20 May 2017, p. 200., doi:10.1016/j.freeradbiomed.2017.10.315.

Tirado, ES;Cortés, AG;Yudasaka, M;Iijima, S;Langad, F;Sedeño, PY;Pingarrón, J;, (2016). Electrochemical immunosensor for the determination of 8-isoprostane aging biomarker using carbon nanohorns-modified disposable electrodes. Journal of Electroanalytical Chemistry.