Tunel stain (Fluorescence)

The In Situ Cell Death Detection Kit, Fluorescein, is a precise, fast, and simple nonradioactive technique to detect and quantify apoptotic cell death at the single-cell level by fluorescence microscopy and quantitative detection by flow cytometry in cells and tissues. Thus, the In Situ Cell Death Detection Kit can be used in many different assay systems.

Examples are:
Detection of individual apoptotic cells in frozen and formalin-fixed tissue sections in basic research
Determination of sensitivity of malignant cells to drug-induced apoptosis in cancer research
Typing of cells undergoing cell death in heterogeneous populations by double staining procedures

Reagent:
In Situ Cell Death Detection Kit, Fluorescein (Roche 11684795910)

Protocol:
1. Dewax and rehydrate tissue section
2. Add proteinase K and incubate 20 minutes under 37°c.
3. Wash slides 2 times with PBS and a 3rd time with PBST (PBS+0.5% Tween20), 5 minutes each.
4. Add 0.1% Triton-PBS and incubate 15 minutes under room temperature.
5. Wash slides 2 times with PBS and a 3rd time with PBST, 5 minutes each.
6.Add prepared TUNEL reagent and incubate 2 hours under 37°c.
7. Wash slides 2 times with PBS and a 3rd time with PBST, 5 minutes each.
8. Add DAPI (1:500) and incubate for 10 minutes under room temperature.
9. Wash slides 3 times with PBS, 5 minutes each.
10. Mount using cover slip resin.