Apoptosis was measured using the Tali™ apoptosis assay kit and propidium iodide (Invitrogen™, Life Technologies, Milan, Italy) as previously reported [24 (link)]. Concisely, on the first day of the assay 1.5 × 105 cells were seeded in a 6-well plate and left to adhere for 16–18 h. The day after seeding, the cells were subjected to UV radiation, with different SPF10-enriched filter combinations, as previously described. At the end of the treatment cells were centrifuged (1500 rpm for 5 min), and resuspended in 100 μL of annexin binding buffer (ABB). After that, 5 μL of Annexin V Alexa Fluor® 488 was added, mixed well, and the solution was incubated in the dark at room temperature for 20 min. Then cells were centrifuged at 1500 rpm, resuspended in 100 μL of ABB, and 1 μL of propidium iodide was added, mixed well, and incubated in the dark at room temperature for 5 min. Samples were analyzed using the Tali® Image-Based cytometer and the percentage of apoptotic nuclei, dead cells, and live cells was determined on the basis of the corresponding fluorescence histogram compared with an untreated control. The results were expressed as fold increase compared with the control.
Tali apoptosis assay kit
Manufactured by Thermo Fisher Scientific
Sourced in Italy
The Tali™ apoptosis assay kit is a fluorescence-based tool for detecting and quantifying apoptosis in cell samples. It provides a reliable and efficient method for evaluating cell viability and apoptosis.
Automatically generated - may contain errors
4 protocols using tali apoptosis assay kit
1
Apoptosis Measurement via Tali Assay
2
Apoptosis Assay of Myometrial and Leiomyoma Cells
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Apoptosis was measured in myometrial and leiomyoma cells using the Tali™ apoptosis assay kit and propidium iodide (Invitrogen™, Life Technologies) according to manufacturer's instructions. According to MTT results, cells were incubated with strawberry extract at 250 μg/ml for 48 h. At the end of incubation, cells were trypsinized, centrifuged (1500 rpm for 5 min), and resuspended in 100 μl of annexin binding buffer (ABB). After that, 5 μl of Annexin V Alexa Fluor® 488 was added, mixed well and the solution was incubated in dark at room temperature for 20 min. Then cells were centrifuged at 1500 rpm, resuspended in 100 μl of ABB and 1 μl of propidium iodide was added, mixed well and incubated in dark at room temperature for 5 min. Samples were analyzed using the Tali® Image-Based cytometer and the percentage of apoptotic nuclei, dead and live cells was determined on the basis of corresponding fluorescence histogram compared with untreated control. Apoptotic cells show green fluorescence, dead cells show red, and live cells show little or no fluorescence.
3
Apoptosis Assay with Annexin V and PI
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Apoptosis was assessed by the Tali™ apoptosis assay kit (Invitrogen™, Life Technologies, Monza, Italy), which uses Annexin V Alexa Fluor™ 488 (Invitrogen™, Life Technologies, Monza, Italy) and propidium iodide (Invitrogen™, Life Technologies, Monza, Italy) to differentiate cells as live, dead, or apoptotic as previously indicated [24 (link)]. Each treatment was performed in three replicates and the final results were reported as a fold increase compared to the control.
4
Apoptosis Assay of Myometrial and Leiomyoma Cells
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Apoptosis was measured using the Tali ® apoptosis assay kit and propidium iodide (Invitrogen™, Life Technologies, Milan, Italy) as previously reported (Islam et al., 2017) . Concisely, on the first day of the assay 1.5 × 10 5 myometrial and leiomyoma cells were seeded in a 6-well plate and left to adhere for 18-20 hr. The day after seeding, the cells were treated for 48 hr with the different strawberry methanolic extracts (250 μg/ml) and with Romina anthocyanin fraction (50 μg/ml). At the end of each treatment, cells were centrifuged and resuspended in 100 μl of annexin binding buffer (ABB). After that, Annexin V Alexa Fluor ® (Thermo Fisher Scientific, Foster, CA) 488 was added, mixed well, and the solution was incubated in the dark at RT. Then, cells were centrifuged, resuspended in ABB, and propidium iodide was added, mixed well, and incubated in the dark at RT. Samples were analyzed using the Tali ® Image-Based Cytometer (Invitrogen™, Life Technologies) and the percentage of apoptotic nuclei, dead cells, and live cells were determined on the basis of the corresponding fluorescence histogram compared with an untreated control. Each treatment was performed three times and the final results were reported as fold increase compared with control.
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