OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation) | STA-325

OxiSelect™ Oxidative RNA Damage ELISA Kit (8-OHG Quantitation)

Introduction:
Free radicals and other reactive species are constantly generated in vivo and cause oxidative damage to
biomolecules, a process held in check only by the existence of multiple antioxidant and repair systems
as well as the replacement of damaged nuclei acids, proteins and lipids. RNA and DNA are probably
the most biologically significant targets of oxidative attack. Recently, oxidative damage to RNA
molecules has been described in several neurological diseases including Alzheimer’s disease,
Parkinson’s disease, Down syndrome, dementia with Lewy bodies, prion disease, subacute sclerosing
panencephalitis, and xeroderma pigmentosum. Among numerous types of RNA oxidative damage, the
formation of 8-hydroxyguanosine (8-OHG) is a ubiquitous marker of oxidative stress.
The OxiSelect™ Oxidative RNA Damage ELISA Kit is a competitive enzyme immunoassay
developed for rapid detection and quantitation of 8-OHG in urine, serum, cerebrospinal fluid or other
cell or tissue RNA samples. The quantity of 8-OHG in unknown sample is determined by comparing
its absorbance with that of a known 8-OHG standard curve. The kit has an 8-OHG detection
sensitivity range of 300 pg/mL to 40 ng/mL. Each kit provides sufficient reagents to perform up to 96
assays, including standard curve and unknown samples.
Assay Principle:
The Oxidative RNA Damage ELISA kit is a competitive ELISA for the quantitative measurement of 8-
OHG. The unknown 8-OHG samples or 8-OHG standards are first added to an 8-OHG/BSA conjugate
preabsorbed EIA plate. After a brief incubation, an anti-8-OHG monoclonal antibody is added,
followed by an HRP conjugated secondary antibody. The 8-OHG content in unknown samples is
determined by comparison with predetermined 8-OHG standard curve.

Kit Components:
Box 1 (shipped at room temperature):
1. 96-well Protein Binding Plate (Part No. 231001): One strip well 96-well plate.
2. Anti-8-OHG Antibody (Part No. 232502): One 15 µL vial of anti-8-OHG.
3. Secondary Antibody, HRP Conjugate (1000X) (Part No. 230003): One 20 µL vial.
4. Assay Diluent (Part No. 310804): One 50 mL bottle.
5. 10X Wash Buffer (Part No. 310806): One 100 mL bottle.
6. Substrate Solution (Part No. 310807): One 12 mL amber bottle.
7. Stop Solution (Part. No. 310808): One 12 mL bottle.
8. 8-OHG Standard (Part No. 232503): One 100 µL vial of 2 μg/mL 8-OHG in 1X PBS, 0.1%
BSA.
Box 2 (shipped on blue ice packs):
1. 8-OHG Conjugate (Part No. 232501): One 20 µL vial of 8-OHG-BSA conjugate at 1.0 mg/mL
in PBS.
Materials Not Supplied:
1. 8-OHG samples such as serum, plasma, urine, cerebrospinal fluid, or RNA extracted from cells
or tissues
2. RNA Extraction Kit
3. Sodium Acetate, pH 5.2
4. Tris Buffer, pH 7.5
5. Nuclease P1, Alkaline Phosphatase
6. 10 kDa molecular weight cutoff (MWCO) centrifuge spin filter (e.g. Amicon Ultra 0.5mL)
7. 10 µL to 1000 µL adjustable single channel micropipettes with disposable tips
8. 50 µL to 300 µL adjustable multichannel micropipette with disposable tips
9. Multichannel micropipette reservoir
10. Microplate reader capable of reading at 450 nm (620 nm as optional reference wave length)
Storage:
Upon receipt, aliquot and store the 8-OHG Standard at -20ºC and the 8-OHG Conjugate at -80ºC to
avoid multiple freeze/thaw cycles. Store all other components at 4ºC.