Hydrogen Peroxide Colorimetric Detection Kit

1. Use plate layout template to aid in proper sample and standard identification. 2. Pipet 50 μL SSA-treated samples, standards, or control into plate wells. 3. Pipet 50 μL Sample Diluent into zero wells. 4. Add 25 μL Fluorescent GSH Detection Substrate to each well using a repeater pipet. 5. Gently tap plate sides to ensure adequate mixing of the reagents. 6. Incubate 15 minutes at room temperature. 7. Read the fluorescent emission at 510 nm with excitation at 370-410 nm. This data will be used to determine Free GSH concentration. 8. Add 25 μL Reaction Mixture to each well using a repeater pipet. 9. Gently tap plate sides to ensure adequate mixing of the reagents. 10. Incubate 15 minutes at room temperature. 11. Read the fluorescence with excitation at 370-410 nm and emission at 510 nm. This data will be used to determine Total GSH concentration.