in situ Cell Death Detection Kit, POD

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Incubate tissue section for 30 min at +37°C with Proteinase K. Add Tunel reaction and incubate for 60 min at +37°C in a humidified atmosphere in the dark	To analyze use an excitation wavelength in the range of 450–500 nm and detection in the range of 515 – 565 nm (green).

Protocol tips
Incubate tissue section for 30 min at +37°C with Proteinase K. Add Tunel reaction and incubate for 60 min at +37°C in a humidified atmosphere in the dark
Downstream tips
To analyze use an excitation wavelength in the range of 450–500 nm and detection in the range of 515 – 565 nm (green).

Publication protocol

The TUNEL assay was performed using the In Situ Cell Death Detection kit, POD (Roche-Applied-Science, Mannheim, Germany), according to the manufacturer's instructions. The paraffin embedding spheres were deparaffinized and rehydrated; then tissues sections were incubated with proteinase K solution (10–20 μg/mL) for 30 min. Tissues were then rinsed twice in PBS and reacted with 50 μL of the TUNEL reaction mixture at room temperature for 60 min in a dark, humidified chamber. Sections were again rinsed in PBS and incubated for 30 min with 50 μL of the Converter-POD (Roche-Applied-Science) and followed by 3-amino-9-ethylcarbazole (AEC). Sections were then counterstained with hematoxylin. As negative controls, corresponding sections were treated in the same way without terminal deoxynucleotidyl transferase.

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Manufacturer protocol

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