Background
Oxidative injury to macromolecules is indicated in a wide range of pathological conditions. Damage is mediated via free radicals that can be created by a range of agents, e.g. xenobiotics, environmental toxins, smoking, radiation, ischaemia-reperfusion injury oxidising agents and normal or disturbed metabolic activity. These free radicals may react with DNA causing reversible and irreversible damage. This can lead to mutation, carcinogenesis, teratogenesis or cell death.
The probe in the OxyDNA Test is specific for 8-oxoguanine. 8-oxoguanine (as part of the oxidized nucleotide 8-oxyguanosine) is formed during free radical damage to DNA and is a sensitive and specific indicator of oxidative DNA damage. 8-oxoguanine is a particularly important biomarker of oxidative DNA damage as it is formed in relatively large quantities and 8-oxoguanine formation can lead to mutations, such as the substitution of thymine for guanine and cytosine for adenine. Previously, 8-oxoguanine was difficult to detect, requiring HPLC analysis, however by utilizing a binding protein with high avidity and specificity for 8-oxoguanine, our OxyDNA Test is a simple, convenient, sensitive fluorescence method for detecting for oxidative DNA damage.
Intended Use
The OxyDNA Test is an in-vitro fluorescent protein binding method for the detection of oxidative damage to DNA in fixed permeabilised cells using the Fluorescent Activated Cell Sorting (FACS) technique. Material of both human and animal origin can be tested.
Principle of the Procedure
The OxyDNA Test utilizes a direct fluorescent protein binding technique. After cells have been fixed and permeabilised, the FITC labeled protein conjugate is added and binds to the 8-oxoguanine moiety present in the 8-oxoguanosine of oxidized DNA. The presence of oxidized DNA is indicated by a green/yellow fluorescence that can be read using the Fluorescence Activated Cell Sorting procedure.
Performance Characteristics
The OxyDNA Test is specific for the 8-oxoguanine moiety of 8-oxoguanosine present in oxidized DNA and shows no measurable cross reactivity with unoxidised guanine / guanosine or other unoxidised nucleotides.
For more information on this product and it's usage see the links to product documents on this page. Alternatively, feel free to contact us.