| 1. Detection Buffer | 5 min |
| 2. Detection Buffer 1 with 0.5% blocking Reagent | 60 min |
| 3. Detection Buffer 1 with 1% BSA, 0.3% Triton | 30–60 min |
| 4. Anti-DIG-AP | 60 min |
| (diluted 1:3K in Buffer 1 with 1% BSA, 0.3% Triton) | |
| 5. Detection Buffer 1 with 0.3% Triton | 10–20 min |
| 6. Detection Buffer 1 with 0.3% Triton | 10–20 min |
| 7. Detection Buffer 1 with 0.3% Triton | 10–20 min |
| 8. Detection Buffer 1 with 0.3% Triton | 10–20 min |
| 9. Detection Buffer | 5 min |
| 10. Detection Buffer | 5 min |
| 11. Color reaction | 1–3 days |
| (under darkness for overnight up to 3 days at room temperature) | |