Description
Description: The Basic Cytotoxicity Test Assay Kit is a single-tube, dual-color assay for determining cell toxicity by flow cytometry. The assay employs a green fluorescent cellular stain, CFSE, to label target cells and the red live/dead viability dye 7-AAD (7-aminoactinomycin D) to identify the killed/dead cells present in the cytotoxicity assay samples. Using this basic cell-mediated cytotoxicit assay, all of the target cells are initially labeled with CFSE. Unstained immune effector cells, cytotoxic drug compounds, or antibodies + complement can then be added and incubated for the desired length of time. Following incubation, 7-AAD dye is added to stain dead cells red. The cells are now ready for analysis. The percentage of cell cytotoxicity is calculated by counting the number of dead target cells (that are stained both green and red) and dividing by the total number of target cells in the sample (all green cells).
Background: Cytolytic activity is an important process by which the immune system eliminates compromised cells in the body. Whether describing antibody- dependent cell-mediated cytotoxicity (ADCC), complement-mediated cytotoxicity, or lymphocyte-mediated cytotoxicity, the activation of the immune system leads to the removal of target cells infected by pathogens or transformed by cancer. Alternatively, cytotoxicity can simply refer to any compound (drug or toxin) that also results in killing of the target cells - intended or otherwise. ICT has developed two easy, reliable, and accurate assays to measure cell-mediated cytotoxicity. Use ICT's Basic Cytotoxicity Test Assay Kit to measure levels of necrotic and membrane-compromised target cells after exposure to effector cells. Use ICT's Total Cytotoxicity Test Assay Kit to further identify apoptotic cells and distinguish apoptosis from necrosis, leading to a more accurate assessment of cell-mediated cytolytic activity. At least 10% more cytolytic activity is often measured when apoptotic cells are also identified. Direct measurement of cytotoxicity rather than using an indirect indicator, such as the release of LDH or ATP enzymes Whole cell assay: no need to lyse cells No radioisotopes: much safer and more cost effective than 51Cr assays