Click-iT™ TUNEL Alexa Fluor™ 488 Imaging Assay

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 2 × 10^4 cells/well were seeded. - Cells were pre-treated with ionizing radiation (5 Gy).	- Cells were fixed with 4 % PFA for 20 min. - Click-iT® reaction is done for 30 min and counter stained for another 30 min at room temperature.

Upstream tips
- 2 × 10^4 cells/well were seeded. - Cells were pre-treated with ionizing radiation (5 Gy).
Protocol tips
- Cells were fixed with 4 % PFA for 20 min. - Click-iT® reaction is done for 30 min and counter stained for another 30 min at room temperature.

Publication protocol

Cell apoptosis were evaluated using Click-iT® TUNEL Assay kit (Invitrogen, USA). Briefly, A172cells were seeded to 96-well plates at the density of 2 × 104 cells well−1 and cultured overnight. Then, cells were treated with ionizing radiation (5 Gy). Following 24-h incubation, cells were fixed with 4 % paraformaldehyde in PBS at room temperature for 20 min. Then the cells were permeabilized with Triton X-100 for 20 min followed incubation with terminal deoxynucleotidyltransferase reaction buffer for at least 10 min at room temperature. TUNEL reaction mixture was added to each well and continued to incubate for 1 h at 37 °C. Following washes with 3 % BSA in PBS for 2 min with gentle shaking, Click-iT reaction mixture was added and incubated for 30 min at room temperature. Afterwards, the cell nuclei were counterstained using Hoechst 33342 for 15 min at room temperature. The positive nuclei were measured by counting the TUNEL-positive cells in eight different, random fields of view each well.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells using Click-iT™ TUNEL Alexa Fluor™ 488 Imaging Assay from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Click-iT™ TUNEL Alexa Fluor™ 488 Imaging Assay below.

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