DeadEnd™ Fluorometric TUNEL System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
1065 cells/well	Transfect for 48h	- When there was a little or poor staining, optimise permeabilization step by adjusting incubation time with permeabilization agent. - When there is a loss of tissue from slides, decrease Proteinase K incubation time.

Upstream tips
1065 cells/well
Protocol tips
Transfect for 48h
Downstream tips
- When there was a little or poor staining, optimise permeabilization step by adjusting incubation time with permeabilization agent. - When there is a loss of tissue from slides, decrease Proteinase K incubation time.

Publication protocol

DNA fragmentation was evaluated by the DeadEndTM Fluorometric TUNEL kit (Promega) according to the manufacturer's directions and flow cytometric quantification of hypodiploid cells (sub-G1 fraction). For quantification of hypodiploid events, 105 cells/well were transfected for 48 h as described above. Staurosporine (STP, Sigma) was used at a concentration of 5 μM as a positive control of apoptosis. Cells were harvested and centrifuged at 600 × g for 5 min. Pellets were suspended in 100% ethanol and stored at −20 °C for 24 h. Cells were then treated with 1 mg/ml RNase A for 1 h at RT. PI was added, and samples were subjected to flow cytometry.

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Papers

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Manufacturer protocol

Download the product protocol from Promega for DeadEnd™ Fluorometric TUNEL System below.

Download manufacturer protocol

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