Live-Dead Cell Fixing Protocol

1. Add 50 ml of DMSO to a vial of Violet Fluorescent Reactive Dye from the LIVE-DEAD Fixable Violet Dead Cell Staining Kit (Cat.No. L34955).
2. Remove any solutions containing sodium azide, Tris buffers or proteins from suspension of cells
3. Adjust cell concentration to 1 x 10⁶/ml.
4. Add 1 ml of amine reactive dye to cell suspension.
5. Mix on Vortex
6. Incubate for 30 mins at RT or on ice.
7. Wash cells in PBS.
8. Add 100 ml of Solution A from Caltag Fix & Perm Kit.
9. Mix on Vortex.
10. Incubate at RT for 15mins.
11. Wash and resuspend in the required volume of PBS.
12. Analyse when required by flow cytometry using the violet 440 nm channel.