Promega
DeadEnd™ Fluorometric TUNEL System
G3250
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This kit was used to detect and quantify the apoptotic cells in a cell population treated with anticancer compounds compared to control cells. The product is simple to use and has high throughput.
For detecting and quantifying the apoptotic cells
Neurobalstoma cells (N2A cells)
Protocol is followed as per the user guide (the promega technical bulletin). In brief, cells were fixed in 4% PFA , 1XPBS washed twice for 5min at RT, cells were permeabilized in 0.2%Tritonx-100(1XPBS) for 5min, 1XPBS wash 5minx2 times @ RT, added and kept with euilibration buffer at RT for 5-10 min, Buffer was removed and the rTdT incubation buffer was added at 37C for 60min in humidified chamber (light sensitive), the reaction was stopped by dipping in 1xSSC (15min RT), washed with deionised water (5minx3times), the coverslip was mounted on slides with vectorshild+DAPI, then it was observed under under a flourescent microscpe.
Step 7 is light sensitive.
In anticancer compound treated cells the number of apoptotic cells are more compared to the control.
None
Simple, high throughput, non-radioactive, used for cells and tissues
High cost
Recommended to use for checking and quantifying the apoptotic cells in various exprerimental conditions (adherent cells).
Product Name: DeadEnd Fluorometric TUNEL SystemSupplier Page