Fluorometric Intracellular Ros Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 0.5 × 105 cells were platted in a 96 well plate.	- 20 μl ROS detection reagent stock solution and 10 ml assay buffer and incubated at 5% CO2, 37°C incubator for 1 hr.

Upstream tips
- 0.5 × 105 cells were platted in a 96 well plate.
Protocol tips
- 20 μl ROS detection reagent stock solution and 10 ml assay buffer and incubated at 5% CO2, 37°C incubator for 1 hr.

Publication protocol

Isolated cardiomyocytes as described above were rapidly transferred into 96‐well flat‐bottom plate (black), 90 μl/well with equivalent amount of 0.5 × 105 cells. Intracellular ROS was determined using a Fluorometric Intracellular ROS Kit (Sigma‐Aldrich). In brief, the master reaction mix (containing 20 μl ROS detection reagent stock solution and 10 ml assay buffer) was added into each well of the cell plate and incubated the plate in a 5% CO2, 37°C incubator for 1 hr. The fluorescence intensity reading at λex = 490/λem = 525 nm was measured by 96‐well plate reader (Thermo Waltham, MA, USA).

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Manufacturer protocol

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