SAT-3 is a stable, highly water-soluble o-Tolidine analog. It is easily oxidized by peroxidase and hydrogen peroxide into a green dye at pH 4-6. While TMBZ requires an organic solvent or detergent to solubilize, SAT-3 can be solubilized with just buffer solution. The SAT-3 solution can be stored at room temperature with protection from light. The maximum wavelength of the dye is 675 nm. Using the Ames test, Nno mutagenicity of SAT-3 was detected. using the Ames test.
2. Add 100-200 μl of the assay solution to each well of a 96-well plate and incubate at 37ºC for 5-30 minutes.
3. Read the O.D. at 670 nm or add 50 μl of 4 M sulfuric acid to each well and read the O.D. at 490 nm instead.