CST, 72782S, 7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit

Protocol
Available protocols: Flow Cytometry (Fixed/Permeabilized)
Background
Cytolytic assays are routinely used to interrogate immune effector cell function. In order to interrogate immune effector cell cytolytic activity in a heterogeneous cell population of effector and target cells, it is imperative to be able to discriminate between effector and target cell populations with distinct phenotypes. As such, this kit is designed to label live target cells with the membrane permeable fluorescent dye CFSE prior to coculture with unlabelled effector cells, thus permitting a clear separation between live effector and target cells. After incubation of live effector and target cells, the DNA intercalating dye 7-AAD is added to label dying target cells with compromised plasma membranes. By using dyes with distinct spectral properties, a clear separation between four cell populations can be obtained using flow cytometry: live target cells, dead target cells, live effector cells, and dead effector cells (1).