In Situ Cell Death Detection Kit, Fluorescein

TUNEL assay cell type - HeLa cells human cervical cancer

Experiment
TUNEL assay cell type - HeLa cells human cervical cancer
Product
In Situ Cell Death Detection Kit, Fluorescein from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Upstream tips
- 2.5 × 10^4 cells per well in 24 well plate were seeded.
Protocol tips
- Cells were stained with anti-TDP-43 (1:150), anti-GFP (1:1000), Alexa 647 anti-rabbit (1:1000) and Alexa 488 anti-chicken (1:1000).

- The enzyme solution (terminal transferase) and label solution were mixed in a volume ratio of 1:9

Publication protocol

Deoxynucleotidyl-transferase-mediated dUTP nick end-labeling (TUNEL) staining was performed using an in situ cell death detection kit (Roche Applied Science, IN). Cells were seeded on poly-D-lysine coated cover slips at a density of 2.5 × 104 cells per well in 24 well plate. After treatment, cells were fixed and stained with anti-TDP-43 (1:150; ProteinTech, Chicago, IL), anti-GFP (1:1000; Invitrogen, Carlsbad, CA), Alexa 647 anti-rabbit (1:1000; Invitrogen, Carlsbad, CA) and Alexa 488 anti-chicken (1:1000; Invitrogen, Carlsbad, CA). The enzyme solution (terminal transferase) and label solution were mixed in a volume ratio of 1:9 to obtain the TUNEL reaction mixture. The cells were incubated with the 50 μl TUNEL reaction mixture at 37°C for 1 hour and then washed three times with PBS. Then the cover slips were mounted on glass slides with antifade reagent (Invitrogen, Carlsbad, CA). Images were obtained with an Olympus Fluoview FV1000 Confocal scanning microscope.

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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

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Papers

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

Paper title
Reducing TDP-43 aggregation does not prevent its cytotoxicity
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Manufacturer protocol

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

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