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. Author manuscript; available in PMC: 2020 May 13.
Published in final edited form as: Methods Mol Biol. 2019;1933:49–65. doi: 10.1007/978-1-4939-9045-0_3
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)