Detecting and Quantifying Apoptotic Cells using DeadEnd™ Fluorometric TUNEL System

Centre for Cellular & Molecular Biology
Project JRF
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Promega

Product Name:

DeadEnd™ Fluorometric TUNEL System

Catalog Number:

G3250

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.

Experimental Design and Results Summary

Application

For detecting and quantifying the apoptotic cells

Starting Material

Neurobalstoma cells (N2A cells)

Protocol Overview

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.

Tips

Step 7 is light sensitive.

Results Summary

In anticancer compound treated cells the number of apoptotic cells are more compared to the control.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Simple, high throughput, non-radioactive, used for cells and tissues

The Bad

High cost

The Bottom Line

Recommended to use for checking and quantifying the apoptotic cells in various exprerimental conditions (adherent cells).

Join the discussion