Image-IT™ LIVE Green Reactive Oxygen Species Detection Kit, for microscopy

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells were pretreated with Sal A for 4 h and then washed before adding 10 µM ATO and incubating for 24 h	- Cells were resuspended in 400 mL of the 25 mM carboxy-H2DCFDA working solution, incubated in the dark for 20–30 min at 37 °C,

Upstream tips
- Cells were pretreated with Sal A for 4 h and then washed before adding 10 µM ATO and incubating for 24 h
Protocol tips
- Cells were resuspended in 400 mL of the 25 mM carboxy-H2DCFDA working solution, incubated in the dark for 20–30 min at 37 °C,

Publication protocol

To explore the effect of Sal A on the intracellular and mitochondrial ROS levels, cells were pretreated with Sal A for 4 h and then washed before adding 10 µM ATO and incubating for 24 h. Then, the cells were harvested and washed with 1× wash buffer. The cells were centrifuged at 400×g for 5 min at room temperature, and the supernatant was discarded. Intracellular ROS levels were measured using an Image-iTTM LIVE Green Reactive Oxygen Species Detection Kit according to the manufacturer’s instructions (Enzo Life Sciences, Farmingdale, NY, USA). The cells were resuspended in 400 mL of the 25 mM carboxy-H2DCFDA working solution, incubated in the dark for 20–30 min at 37 °C, and then analyzed via flow cytometry. The mitochondrial ROS levels were measured using MitoSOXTM Red mitochondrial superoxide indicator (Invitrogen, Molecular Probes). After treatment, the cells were suspended in warm PBS and incubated with 5 µM MitoSOX reagent for 10 min at 37°C in the dark. The cells were then washed three times with PBS, and the mitochondrial ROS levels were analyzed using flow cytometry

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

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing ROS assay cell type - H9c2 rat cardiomyocytes using Image-IT™ LIVE Green Reactive Oxygen Species Detection Kit, for microscopy from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Image-IT™ LIVE Green Reactive Oxygen Species Detection Kit, for microscopy below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing ROS assay cell type - H9c2 rat cardiomyocytes using Image-IT™ LIVE Green Reactive Oxygen Species Detection Kit, for microscopy from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.