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Assay Kits & ELISAs

Cell-mediated Cytotoxicity Assay: Total Cytotoxicity Test

Product Sizes
125 tests
971-125TESTS
About this Product
SKU:
971
Application:
Flow Cytometry
Extra Details:
A single-tube, tri-color fluorescent assay for quantitative assessment of apoptosis and necrosis. CFSE (green) labels target cells, 7-AAD (red) labels dead cells, and SR-FLICA® (orange-red) labels active caspases. Analyze by flow cytometry.
Immunogen:
Cell-mediated cytotoxicity, poly caspases
Physical State:
Cell Culture
Shipping Conditions:
Blue Ice
Storage Conditions:
2-8[o]C
Type:
Assay Kits: Cell Viability
Figure 1. Quantification of 4 populations in each sample
Figure 10. Control D contains healthy target cells labeled with CFSE and SR-FLICA® that were not induced to undergo apoptosis. Control E contains target cells labeled with CFSE and SR-FLICA® that were induced to undergo apoptosis. Apoptotic target cells stained green with CFSE and orange-red with SR-FLICA® (Control E) exhibit high levels of staining with SR-FLICA (SR-VAD-FMK) and migrate to the right compared with non-induced cells (Control D). See protocol.
Figure 11. Control F contains live target cells stained with CFSE and 7-AAD. Control G contains killed target cells stained with CFSE and 7-AAD. Run Controls F (Figure 11A) and G (Figure 11B) to compensate CFSE (FL-1) vs. 7-AAD (FL-3). Killed target cells stained green with CFSE and red with 7-AAD (Control G) migrate up the plot compared to live cells (Control F). Create a gate (R3) on the green target cell population and rerun Control A (Figure 12). See protocol for details.
Figure 12. Control A contains CFSE stained target cells and effector cells (E:T). Create a dot plot of SR-FLICA® (FL-2) vs. 7-AAD (FL-3) gated off R3. CFSE stained target cells appear in the lower left quadrant of this plot. Since Control A has not been stained with either SR-FLICA (SR-VAD-FMK) or 7-AAD, there should be very few cells outside this region. See protocol for details.
Figure 13. Control H contains non-induced CFSE stained target cells labeled with SR-FLICA and 7-AAD. Run Control H in SR-FLICA® (FL-2) vs. 7-AAD (FL-3). This control reveals background levels of spontaneous death and apoptosis in the non-induced cells without the influence of the effector cells. See the protocol for details.