5-Carboxyfluorescein diacetate N-succinimidyl ester

5-Carboxyfluorescein diacetate N-succinimidyl ester is a cell permeable dye (Ex=492 nm, Em=517 nm). 5-Carboxyfluorescein diacetate N-succinimidyl ester can label cells by covalently binding to intracellular molecules. 5-Carboxyfluorescein diacetate N-succinimidyl ester is used to track lymphocyte migration and proliferation^[1]. In Vitro:Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[1].
1. Dilute the stock solution in PBS (to give a 50 μM solution).
2. Add 110 μL of this solution per milliliter of cells (to give a final concentration of 5 μM) and mix rapidly.
3. After 5 min at room temperature, add 10 vol of PBS containing 5% FBS.
4. Centrifuge cells 5 min at 300 ×g, 20 ℃.
5. Remove the supernatant, and wash threetimes, each time by resuspending in 10 vol PBS containing 5% FBS.
6. Centrifuge cells 5 min at 300 × g, 20 ℃, and remove the supernatant.
Note: Labeling occurs rapidly, and it is essential that the tracker is dispersed as evenly and quickly as possible so that cells are uniformly labeled.