OxiSelect Protein Radical ELISA Kit

OxiSelect Protein Radical ELISA Kit

Cat. Num. AAA169163

• Synonyms: OxiSelect Protein Radical; OxiSelect Protein Radical ELISA Kit; OxiSelect Protein Radical elisa kit

• Preparation and Storage: Upon receipt, aliquot and store the DMPO Nitrone Adduct HSA and Reduced BSA Standard at -20 degree C to avoid multiple freeze/thaw cycles. Store all other components at 4 degree C.

Typical Testing Data/Standard Curve (for reference only)

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Related Product Information for OxiSelect Protein Radical elisa kit

Principle of the Assay: DMPO Nitrone Adduct-HSA standard or DMPO adduct protein samples are adsorbed onto a 96-well plate for 2 hrs at 37 degree C or 4 degree C overnight. The DMPO protein adducts present in the sample are probed with an anti-DMPO Nitrone Adduct antibody, followed by an HRP conjugated secondary antibody. The protein radical content in the unknown sample is determined by comparing with a standard curve that is prepared from predetermined DMPO Nitrone Adduct-HSA standards.

Background/Introduction: Reactive nitrogen species (RNS) such as peroxynitrite (ONOO-) and reactive oxygen species (ROS) such as peroxyl radicals (ROO·), hydroxyl radicals (OH·), or superoxide radical anions have been implicated in protein radical formation through the removal of an electron or hydrogen atom. These protein radicals can lead to internal crosslinking of the polypeptide chain, cause protein backbone cleavage, and form protein peroxides as well as peroxyl radicals. The formation of protein radicals has been linked to various disorders including amyotrophic lateral sclerosis, aging, Huntington's Disease, and Alzheimer's disease. Radical formation results in an unstable molecule that contains an atom with one unpaired electron in its outer orbital, giving the atom unique paramagnetic properties. These properties allow for the use of the electron spin resonance (ESR) technique to detect various forms of biological radicals. Since radicals are very unstable, an ESR spin trapping technique was developed to increase detection sensitivity. ESR spin trapping involves a reaction between the reactive protein radical and a diamagnetic molecule (such as 3,4-dihydro-2,2-dimethyl-2H-pyrioole 1-oxide, or DMPO) also known as a spin trap. This reaction results in a more stable DMPO radical adduct that can be detected by ESR. More recently, an "immuno-spin trapping" technique has been used to detect radical formation (Figure 1). It has been demonstrated that the aforementioned DMPO radical adduct decays into a DMPO nitrone adduct due to oxidation. Antibodies raised against the stable DMPO nitrone adduct recognize DMPO bound to protein, and can be used to demonstrate protein radical formation in an ELISA or western blot format. The OxiSelect™ Protein Radical ELISA Kit is an immunoassay developed for rapid detection and quantitation of protein radicals. The relative amount of protein radicals within a protein sample is determined by comparing its absorbance with that of a known DMPO Nitrone Adduct Human Serum Albumin (HSA) standard curve. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown protein samples.

Product Categories/Family for OxiSelect Protein Radical elisa kit

Oxidative Stress/Damage; Protein Oxidation/Nitration; Protein Radical ELISA

References

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