Product nameCytotoxicity Assay Kit (CFSE, 7-AAD)
Cytolytic activity is an important process for eliminating intracellular pathogens and cancer cells. Cytotoxicity Assay Kit (CFSE, 7-AAD) (ab270780) allows you to assess cytolytic activity in cell culture.
Older methods to assess NK cytolytic activity include measuring the release of lactate dehydrogenase, and more commonly, the release of radioactive 51Cr from lysed target cells. Unfortunately, these techniques have several drawbacks. Traditional enzyme-release assays are often skewed by the large number of necrotic effector cells. Problems associated with 51Cr release methods include high spontaneous leakage resulting in high backgrounds, high cost, short half-life, and the health risks due to exposure to radioactive material.
Flow cytometric assays have been developed to overcome some of the difficulties associated with older assays like lactate dehydrogenase and 51Cr release assays. One such early version involved the detection of NK cytotoxicity activity by staining target cells with the green fluorescent dye, F-18, in combination with the DNA intercalating dye, propidium iodide . Since then, a red fluorescent membrane dye, PKH-26, has been used in preference to F-18, and in combination with the viability probe, TO-PRO-3 iodide4-7. However, despite correlations of greater than 95% when compared with the 51Cr release assay, the PKH-26 method is problematic. It is difficult to use at a constant concentration, leading to unreliable staining, and the staining procedure requires multiple steps, often decreasing the viability of the target cells.
Cytotoxicity Assay Kit (CFSE, 7-AAD) includes two fluorescent reagents: CFSE and 7-AAD. CFSE, a green fluorescing cellular stain, is utilized to identify the target cell population. The unstained effector cells are added and incubated with the target cells. 7-AAD, a red fluorescing live/dead stain, is then added to stain all necrotic cells red by binding to the DNA of membrane-compromised cells. The target cell and effector cell populations can then be easily distinguished, and flow cytometry easily identifies live and necrotic cells.
Tested applicationsSuitable for: Flow Cytmore details
Storage instructionsPlease refer to protocols.
Components 125 tests 250 tests 10X Assay Buffer 1 x 30ml 1 x 60ml 7-AAD, red live/dead stain, 125 tests 1 vial 2 vials CFSE, green cellular stain, 250 tests 1 vial 1 vial
Our Abpromise guarantee covers the use of ab270780 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent concentration.|
ab270780 has not yet been referenced specifically in any publications.