TO-PRO-3 (iodide)

TO-PRO-3 iodide is a cell-impermeant nucleic acid stain. TO-PRO-3 iodide is useful as a nuclear counterstain and dead cell indicator (Ex/Em = 642/661 nm). In Vitro:Instructions (The following is our recommended protocol. This protocol is for guidance only and should be modified according to your specific needs).

1. Preparation of Working Solution
Dilute the stock solution with pre-warmed serum-free cell culture medium or PBS to prepare a working solution of 1-5 μM.
Note: Please adjust the TO-PRO-3 working solution concentration according to actual conditions, and prepare it fresh before use.

2. Staining of Fixed Cells
2.1 Centrifuge to collect cells, and wash twice with PBS for 5 minutes each time.
2.2 Add sufficient staining solution to cover the cells, and incubate at room temperature in the dark for 15-30 minutes.
2.3 Centrifuge at 400 g for 3-4 minutes, and discard the supernatant.
2.4 Wash the cells twice with PBS for 5 minutes each time.
2.5 Resuspend the cells in 1 mL of serum-free culture medium or PBS, and observe using a fluorescence microscope or flow cytometer (Ex/Em = 642/661 nm).