APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cell were pretreated with 50 μM, 75 μM, and 100 μM of honokiol for 12, 24, 48, or 72 hours.	- Cells were fixed with 1% (w/v) paraformaldehyde in PBS and place on ice for 15 minutes	- If signal intensity is low increase the incubation time for the DNA-labeling reaction

Upstream tips
- Cell were pretreated with 50 μM, 75 μM, and 100 μM of honokiol for 12, 24, 48, or 72 hours.
Protocol tips
- Cells were fixed with 1% (w/v) paraformaldehyde in PBS and place on ice for 15 minutes
Downstream tips
- If signal intensity is low increase the incubation time for the DNA-labeling reaction

Publication protocol

The DNA fragmentation in SKMEL-2 and UACC-62 cells was quantified using an Apo-BrdU TUNEL assay kit (Molecular Probes, Eugene, OR, USA) per the manufacturer's protocol as reported earlier [17]. Cells were treated with control media, 50 μM, 75 μM, and 100 μM of honokiol for 12, 24, 48, or 72 hours and harvested and fixed shortly after the completion of treatment. DNA nicks in fixed cells were labeled with BrdU; then samples were incubated with Alexa Fluor 488-labeled anti-BrdU antibody. The cells were analyzed using flow cytometry. Both positive and negative controls were run with each assay. The experiment was repeated four times.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU below.

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