Apoptosis Kits Protocols-FAQS

How do I determine the % of cell death (from my treatment) using this kit? Determine the number of cells that are dual stained green and red. These are your dead (necrotic) cells. Divide this number by the total number of green cells and multiply by 100 to obtain the percentage of your target cells that were killed by the experimental treatment.

What controls are necessary to run this kit? It is important to have a positive (killed target cells) control for a cytotoxicity assay. Use any method you desire to kill your CFSE-labeled target cells (eg. freeze/thawing, treatment with alcohol or staurosporine or camptothecin etc.). These cells can be stained with 7-AAD to facilitate gating of the 7-AAD positive cell population. In addition, it is important to have a CFSE-only stained sample of normal-healthy target cells to facilitate flow cytometer gating/setup. This healthy CFSE stained control should be stained with 7-AAD to subsequently assess the degree of baseline spontaneous cell death present in the normal cell population. . Moreover, it is always a good idea to have a sample of unstained cells to determine the level of auto-fluorescence in the assay and to set up your forward versus side-scatter dot plot on flow cytometer instrument.

What advantages does this Basic Cytotoxicity Kit offer over older more traditional forms of cell-cytotoxicity assessment?  Neuromics's Basic Cytotoxicity Test Kit does not utilize radioactivity, nor rely on the release of intracellular enzymes for detection of membrane compromised, dead or dying cells. ICT's assay kit allows for easy quantitation and visualization of results using flow cytometry and analysis software.