|Summary of procedure:||
The TBARs assay kit measures malondialdehyde (MDA), a reactive compound formed from lipid peroxides that are generated under conditions of oxidative stress. Oxidative modification of lipids occurs with aging and various diseases, and increased oxidative stress is associated with diabetes and its complications. Cellular exposure to oxidative stress gives rise to highly reactive and unstable lipid hydroperoxides. Decomposition of unstable peroxides derived from polyunsaturated fatty acids leads to the MDA formation. MDA reacts with thiobarbituric acid (TBA) to form an adduct that can be measured spectrophotometrically or fluorometrically, the latter providing greater sensitivity. Determination of TBARs in biological samples has become the preferred method for assessing lipid peroxidation and oxidative stress. Exocell’s TBARs assay can be used with a spectrum of biological samples including body fluids, tissue and cell specimens.
To complete the assay, standard or sample is reacted with thiobarbituric acid and heated in the presence of acetic acid. After cooling and clarification by centrifugation, aliquots are placed into microtiter wells of a black plate and read in a fluorometer at excitation 484 nm, emission 530 nm, or into a plastic plate for reading at 530-540 nm in a spectrophotometer. MDA concentration in samples is determined from the standard curve.
|Assay Range:||0-30 nmole fluorometrically
0-300 nmole spectrophotometrically
|Precision:||Intra- and inter-assay precision <8%|