7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit (#72782) Datasheet Without Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:

FC-FP

Product Information

Storage

Kit will arrive packaged as a -20°C kit. For best results, remove 7-AAD Viability Dye and Cell-Based Assay Buffer Tablet and store at 4ºC and RT, respectively. CFSE Stock Solution should be stored at -20ºC. Product is stable for at least 12 months from date of receipt when stored as recommended.

Product Description

The 7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit employs CFSE (carboxyfluorescein succinimidyl ester) to label target cells within the mixed cell population and 7-AAD (7-aminoactinomycin D) to label dead cells. This kit provides an improvement over the traditional chromium-51 (⁵¹ Cr) release assay to assess cell-mediated cytotoxicity. CFSE labeling is more sensitive, does not employ radioisotopes, and cytolysis can be assessed at the single-cell level using flow cytometry.

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).

Kim, G.G. et al. (2007) J Immunol Methods 325, 51-66.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

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