ApopTag Fluorescein in Situ Apoptosis Detection Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 1 x 10^5 cells/well	Treat cells with simvastatin, 10nM docetaxel or a combination of both for 24h. Fix cells in 2% PFA at 4°C for 30 min

Upstream tips
Seed 1 x 10^5 cells/well
Protocol tips
Treat cells with simvastatin, 10nM docetaxel or a combination of both for 24h. Fix cells in 2% PFA at 4°C for 30 min

Publication protocol

The TUNEL assay for in situ detection of apoptosis was performed by using the ApopTag® Fluorescein In Situ Apoptosis detection kit (Millipore, MA) according to the manufacturer’s instructions. Cells were plated in 24-well flat bottom plates at a density of 1 x 105 cells/well and treated with 25 μM simvastatin, 10nM docetaxel or a combination of both for 24h. Following treatments, cells were fixed in 2% paraformaldehyde at 4°C for 30 min. Fixed cells were then permeabilized in 0.1% Triton X-100 and labeled with fluorescein 12-dUTP using terminal deoxynucleotidyl transferase. Nuclei were counterstained with DAPI. Frozen nude mouse prostate tumor (PC3) xenograft sections were also processed accordingly. Cells/tissue sections were analyzed for apoptotic cells with localized green fluorescence using an inverted fluorescence microscope (Zeiss Axiovert100M, Carl Zeiss, Germany).

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Manufacturer protocol

Download the product protocol from Millipore for ApopTag Fluorescein in Situ Apoptosis Detection Kit below.

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