Fluorometric Intracellular Ros Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Reconstitute ROS Detection Reagent with 40 mL of DMSO to generate the 500´ ROS Detection Reagent Stock Solution.	- The Master Reaction Mix is enough for one plate. The Master Reaction Mix is best used within 2 hours.

Upstream tips
- Reconstitute ROS Detection Reagent with 40 mL of DMSO to generate the 500´ ROS Detection Reagent Stock Solution.
Protocol tips
- The Master Reaction Mix is enough for one plate. The Master Reaction Mix is best used within 2 hours.

Publication protocol

The intracellular level of ROS was measured with a fluorometric intracellular ROS kit (Sigma Aldrich) according to manufacturer’s instructions. The fluorescence intensity was detected at lex = 650/lem = 675 nm using a fluorescence microplate reader (Reader Synergy HT, BioTek). The intracellular level of NO production was determined by the Griess Reagent System (Promega) according to manufacturer’s instructions. The absorbance was measured at 540 nm using a microplate reader.

The level of GSH was determined with a glutathione assay kit (Sigma Aldrich) according to manufacturer’s instructions. The fluorescence intensity was measured by a fluorimeter plate reader set at an excitation wavelength of 390 nm and emission wavelength of 478 nm.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Fluorometric Intracellular Ros Kit below.

Download manufacturer protocol

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