| Assay Method Information | |
| | SOD Assay (Fluorimetric Analysis) |
| Description: | SOD activity of the purified samples was determined by the SOD assay kit using (2-(4-iodophenyl)-3-(4nitrophenyl)-5-(2,4-disulphenyl)-2H-tetrazolium, monosodium salt) [WST] which is a colorimetric assay that monitors the rate of inhibition of WST to a water soluble formazan dye. Purified SOD from bovine brain (20 μl) was incubated (37°C, 20 min) with WST working solution (200 μl) and distilled water (20 μl) according to the Sigma Information Bulletin, 19160 after which SOD activity was determined from the absorbance at 450 nm. Spectrofluorimetry was used to analyse the interaction of amyloid peptides [Aβ25-37, Aβ29-33 and Aβ1-40] with SOD. The excitation wavelength was fixed at 295 nm, the wavelength at which tryptophan absorbs, and the emission wavelength was at 482 nm. The change in fluorescence of the solution was monitored as increasing concentrations of amyloid peptides (0-40 μM) were added to a reaction mixture of SOD (5.0 μl) in Tris-HCl buffer (pH 8.0, 50 mM) in a final volume of 300 μl. |
| Affinity data for this assay | |
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