Biotinylated tyramine (BT) solution

Reagent	Final concentration	Amount
Biotin-NHS solution	n/a	36.5 μL
Tyramine hydrochloride solution	n/a	36.5 μL
2-Aminoethanol	n/a	7.3 μL
Total	n/a	80.3 μL

For a detailed procedure of preparation of BT solution, refer to Okamoto et al. (2021).

Mix well biotin–NHS solution and tyramine hydrochloride solution and incubate the mixture for 12–24 h at 20°C–25°C with rotation and protection from light.

Add 7.3 μL of monoethanolamine to the mixture, and incubate it for 4 h at 20°C–25°C with rotation and protection from light.

Complete reaction between biotin-NHS and tyramine hydrochloride yields 128 mM of BT in the solution.

The solution can be stored at −80°C for 3 years.