OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 1×10^5 cells/ml cells were platted for assay	- Treated with 1X solution of DCFH-DA reagent at 37C for 1h

Upstream tips
- 1×10^5 cells/ml cells were platted for assay
Protocol tips
- Treated with 1X solution of DCFH-DA reagent at 37C for 1h

Publication protocol

The levels of reactive oxygen species (ROS) were determined quantitatively and qualitatively by 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA) using the OxiSelect™ kit from Cell Bio Labs (San Diego, CA). For quantitative analysis, cells were separated and adjusted to 1×105 cells/ml. Aliquots from each cell suspension (100 μl) were then gently mixed with a 1X solution of DCFH-DA reagent (100 μl) and incubated at 37°C for 1h. Following this incubation period, cells were lysed according to the manufacturer’s specifications and the reading from each cell suspension was measured with a florometric plate reader (Spectramax, Gemini EM) at excitation and emission wavelengths of 480 and 530 nm, respectively. Qualitative measurement of dye uptake by cells was performed according to the manufacturer’s protocol. All images were captured using an EVOSfl fluorescence microscope equipped with a GFP filter (470 nm excitation, 525 nm emission).

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Manufacturer protocol

Download the product protocol from Cell Biolabs for OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) below.

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