ROS-ID® Total ROS/Superoxide detection kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Cells with treated with oxidative stress detection reagent and superoxide detection reagent for 30 minutes before CNT solution. - After 60 minutes of Pyocyanin treatement, cells were washed, harvested and treated with trypsinethylenediaminetetraacetic acid. - Cells were resuspended in 0.3 mL 1× wash buffer with 10% FBS and passed through a nylon mesh; then assessed by FACS.

Protocol tips
- Cells with treated with oxidative stress detection reagent and superoxide detection reagent for 30 minutes before CNT solution. - After 60 minutes of Pyocyanin treatement, cells were washed, harvested and treated with trypsinethylenediaminetetraacetic acid. - Cells were resuspended in 0.3 mL 1× wash buffer with 10% FBS and passed through a nylon mesh; then assessed by FACS.

Publication protocol

Total ROS/superoxide production in cells exposed to CNTs was determined using a total ROS/superoxide detection kit (Enzo Life Sciences, Inc., Farmingdale, NY, USA) according to the manufacturer’s instructions. After the cells had adhered for 24 hours in 12-well plates, they were pretreated with oxidative stress detection reagent and superoxide detection reagent for 30 minutes before CNT solution (1 μg/mL in BEAS-2B and 10 μg/mL in MESO-1 cells) was added. Pyocyanin (100 μM) was used to induce ROS production. After 60 minutes, the cells were washed once with 1× wash buffer and harvested with trypsin–ethylenediaminetetraacetic acid. Cells were resuspended in 0.3 mL 1× wash buffer with 10% FBS and passed through a nylon mesh; then, they were subjected to flow cytometry (FACSCalibur™; BD Biosciences, San Jose, CA, USA) using the FL1 and FL2 channels for oxidative stress detection reagent and superoxide detection reagent signals, respectively, until 10,000 cells were recorded. Cell suspensions were assayed in triplicate for each treatment condition.

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 - BEAS-2B human bronchial epithelial cell line using ROS-ID® Total ROS/Superoxide detection kit from Enzo Life Sciences.

Manufacturer protocol

Download the product protocol from Enzo Life Sciences for ROS-ID® Total ROS/Superoxide detection kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing ROS assay cell type - BEAS-2B human bronchial epithelial cell line using ROS-ID® Total ROS/Superoxide detection kit from Enzo Life Sciences. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.