ApopTag Fluorescein in Situ Apoptosis Detection Kit

TUNEL assay cell type - PC-3 human prostate cancer

Experiment
TUNEL assay cell type - PC-3 human prostate cancer
Product
ApopTag Fluorescein in Situ Apoptosis Detection Kit from Millipore
Manufacturer
Millipore

Protocol tips

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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Discussion

Discussion

4 years ago

Author: Ercole Udinesi Italy

Are 10 week old samples in formalin still usable?

I have left my tumor samples in formalin for an extended period of time (around 10 weeks). Do you think I will be able to use them for TUNEL assay and get results? Thank you for your help in advance

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Papers

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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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