Note: This protocol is intended for use with the specific products mentioned within it. Substituting different products is not recommended.
The Cell Proliferation Dyes (CPDs) Invitrogen™ eFluor™ 670, CPD Invitrogen™ eFluor™ 450, and 5-(and 6)-Carboxyfluorescein diacetate succinimidyl ester (CFSE) are fluorescent dyes that can be used to monitor individual cell divisions. These dyes may also be used for tracking cells in vivo for up to several weeks in non-dividing cells. The dyes covalently react with cellular proteins. As cells divide, the dyes are divided equally between daughter cells, resulting in successive halving of the fluorescence intensity of the dye. Six to eight generations may be visualized, depending on which dye is used.
Cells labeled with proliferation dyes may be fixed and permeabilized for analysis of intracellular targets using standard formaldehyde containing fixatives and saponin-based permeabilization buffers, such as the Foxp3/Transcription Factor Staining Buffer Set (cat. no. 00-5523) or the Intracellular Fixation & Permeabilization Buffer Set (cat. no. 88-8824)