Protocol: L-amino acid oxidase activity measurement/LAO活性測定のプロトコル
Use 96 well flat-bottomed micro titer plate
Step 1. Add 5 micro-L of 100 μM L-amino acid solution in 50 mM HCl to well
Step 2. Add 20 micro-L of 5 M NaCl solution
Step 3. Add 50 micro-L of 0.25 mg/mL o-phenylenediamine contained 0.1 unit/mL horseradish peroxidase in 1 M Tris-HCl buffer (pH 7.4)
Step 4. Mix 25 micro-L of sample solution or hydrogen peroxide standard (0~1.0 mM serial dilution series)
Step 5. Incubate at 37 degree C for 30~150 min
Step 6. Terminate reaction by adding 100 micro-L of 1 M H2SO4
Step 7. Measure absorbance at 492 nm using microplate reader