TUNEL Assay Kit - BrdU-Red

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Grow cells without any growth factors for 24 h. Treat cells with Cur or SLCP (25 μM) for 24–72 h	fix cells with 4% paraformaldehyde for 15 min,	View slides as soon as possible. Analyze cells within 3 hours of staining.

Upstream tips
Grow cells without any growth factors for 24 h. Treat cells with Cur or SLCP (25 μM) for 24–72 h
Protocol tips
fix cells with 4% paraformaldehyde for 15 min,
Downstream tips
View slides as soon as possible. Analyze cells within 3 hours of staining.

Publication protocol

The TUNEL staining was performed as per manufacturer's instructions [28, 35]. Briefly, U-87MG cells were grown on coverslips in EMEM, without any growth factors for 24 h and then they were treated with Cur or SLCP (25 μM) for 24–72 h. Following treatment, the cells were fixed with 4% paraformaldehyde for 15 min, and then TUNEL staining was performed [28, 35]. Finally, the cells were counterstained with Hoechst 3342 for 5 min at room temperature in the dark and washed thoroughly with distilled water, after which they were mounted on a glass slide with antifading medium. The cells were observed under a fluorescent microscope (Leica, Germany), using appropriate filters (ex/em: 488/576). The red fluorescent signal indicated TUNEL-positive cells. The number of total cells and that of TUNEL-positive cells were counted and expressed as a percentage of the total cell count. Almost two thousand total cells were counted in each group of randomly selected microscopic fields from three independent experiments to obtain a mean value.

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Manufacturer protocol

Download the product protocol from Abcam for TUNEL Assay Kit - BrdU-Red below.

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