Cytotoxicity Dye Kit (CFSE, 7-ADD)
The Cytotoxicity Dye Kit (CFSE, 7-AAD) is a kit designed to determine the cytotoxicity profile of effector cells towards target cells of interest. It consists of CFSE (carboxyfluorescein succinimidyl ester), used to identify target cells in a mixed cell population of target cells and PBMC, and 7-AAD (7-aminoactinomycin D) to label dead cells. CFSE and 7-AAD labeling enables cell cytotoxicity detection at the single-cell level using flow cytometry. The kit also contains Assay Diluent.
- Effector cells of interest.
- Target cells of interest.
- U-bottom 96 well plate for cell culture.
- T75 cell culture flask.
| Catalog # | Name | Amount | Storage |
| 82177 | 500x CFSE | 1 vial | -20°C |
| 82178 | 1000x 7-ADD | 1 vial | 4°C |
| 82184 | Assay Diluent | 25 ml | 4°C |
Lymphocyte-mediated cytotoxicity is a form of cellular immunity targeting intracellular pathogens, including viruses and certain bacteria and parasites. The most popular in vitro methods to monitor lymphocyte-mediated cytotoxicity of target cells are cell-mediated cytotoxicity assays such as ADCC (antibody-dependent cellular cytotoxicity) and TDCC (T cell-dependent cellular cytotoxicity), in which immune effector cells and target cells are co-cultured. To analyze immune effector cell cytolytic activity in such heterogeneous cell populations it is essential to be able to discriminate between effector and target cell populations, with distinct phenotypes. The use of a label prior to co-culture with effector cells, like the membrane-permeable fluorescent dye CSFE (carboxyfluorescein succinimidyl ester), able to stain live target cells permits a clear distinction between live effector and target cells. After incubation with target cells, the DNA intercalating dye 7-ADD can be added to label dying target cells. These two dyes have distinct spectral properties and allow four cell populations to be discriminated by flow cytometry: live target cells (CSFE+7AAD-), dead target cells (CSFE+7AAD+), live effector cells (CSFE-7AAD-), and dead effector cells (CSFE-7AAD+). Cytotoxicity assays are crucial to understand the potency of CAR (chimeric antigen receptor) T and NK cells, and antibody-based immunotherapies.
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