In Situ Cell Death Detection Kit, Fluorescein

TUNEL assay cell type - PC-3 human prostate cancer

Experiment
TUNEL assay cell type - PC-3 human prostate cancer
Product
In Situ Cell Death Detection Kit, Fluorescein from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Upstream tips
Seed 10,000 cells in 96-well plate.

Fix sample in 4% formaldehyde for 1 h before freezing.
Protocol tips
Fix sample in 4% formaldehyde for 5 min before using sample.

Block samples in 10% donkey serum in 0.1% Triton-X for 30 min at room temperature.

Incubate primary antibody overnight at 4 °C and with secondary antibodies for 1 h at room temperature

Publication protocol

The apoptotic cells were quantified for DNA fragmentation, by using an In-situ Cell Death Detection kit, Fluorescein (Roche Diagnostics GmbH, Mannheim, Germany), according to the manufacturer’s protocol. In brief, excised tumor tissues were fixed in 4% paraformaldehyde, dehydrated and embedded in paraffin. Paraffin-embedded tissues were cut into 5-μm thick sections. The sections were deparaffinized and rehydrated according to standard protocol. The tissue sections were then antigen retrieved at 0.1 M citrate buffer for ∼5 min in a pretreatment module (pH 6.0) and then rinsed twice in PBS. The slides were then treated with 50 μl of ‘TUNEL reaction mixture’ and incubated at 37 °C for 60 min The slides were then rinsed 3 × with PBS. Negative and positive slides were included. For negative slides, only 50 μl of ‘Label solution’ (without terminal transferase enzyme) was used and for positive slides, the slides were treated with DNase I before proceeding with treating with ‘TUNEL reaction mixture’. For microscopic image capture, four high-power fields were randomly selected and the apoptotic index was calculated as equal to number of positive cells/total number of cells × 100%.

Full paper   Login or join for free to view the full paper.

Reviews

In Situ Cell Death Detection Kit, Fluorescein from Sigma-Aldrich has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Discussion

5 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

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing TUNEL assay cell type - PC-3 human prostate cancer using In Situ Cell Death Detection Kit, Fluorescein from Sigma-Aldrich.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Sigma-Aldrich for In Situ Cell Death Detection Kit, Fluorescein below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing TUNEL assay cell type - PC-3 human prostate cancer using In Situ Cell Death Detection Kit, Fluorescein from Sigma-Aldrich. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms