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Cat. No.: ROSCH-200 (for 20-200 samples)

Cat. No.: ROSCH-1k (for 100-1000 samples)

Description

Reactive Oxygen Species Assay Kit with CM-H2DCFDA, also known as the ROS Assay Kit with CM-H2DCFDA, is designed to detect reactive oxygen species (ROS) using the fluorescent probe CM-H2DCFDA. This kit is an upgraded version of the Reactive Oxygen Species Assay Kit.

This kit utilizes the fluorescent probe DCFH-DA (H2DCFDA) for ROS detection, whereas CM-H2DCFDA is a chloromethyl derivative of H2DCFDA (also known as CM-DCFH-DA). CM-H2DCFDA has better retention in live cells compared to H2DCFDA, resulting in stronger fluorescence signals and making it more suitable for relatively longer studies of intracellular ROS.

CM-H2DCFDA itself is non-fluorescent and can freely pass through cell membranes. Once inside the cell, it is hydrolyzed by intracellular esterases to produce CM-H2DCF (also known as CM-DCFH). CM-H2DCF cannot cross cell membranes, ensuring that the probe is easily loaded into cells. The chloromethyl group of CM-H2DCF reacts with intracellular glutathione or other thiols, providing better intracellular retention. Subsequent oxidation of non-fluorescent CM-H2DCF by ROS generates fluorescent CM-DCF. By detecting the fluorescence of CM-DCF, the level of intracellular ROS can be determined.

This kit includes Rosup, a positive control reagent for ROS detection. Rosup is a compound mixture with a concentration of 50mg/ml.

The kit has a low background, high sensitivity, wide linear range, and is easy to use. It comes in small and medium packaging. For a 1 ml detection system per well in a 6-well plate, it can perform 20 and 100 tests, respectively. For a 100 µl detection system per well in a 96-well plate, it can perform 200 and 1000 tests, respectively. When used with a flow cytometer, with a detection system volume of 0.5 ml per sample, it can perform 40 and 200 tests, respectively.

Components

• CM-H2DCFDA (5mM)
• Positive control

Storage

Store at -20ºC, valid for one year. CM-H2DCFDA (5mM) must be stored away from light.

Precautions

• After loading the probe, be sure to wash off any residual probe that did not enter the cells to avoid high background. Once the probe is loaded and residual probe is washed off, you can perform excitation and emission wavelength scans to confirm proper probe loading. Minimize the time between probe loading and measurement (excluding stimulation time) to reduce potential errors.
• For fluorescence plate reader detection, use black or white plates suitable for fluorescence detection. We recommend Ultra-Low Attachment Black 96-Well Plates with Clear Flat Bottom.
• This product is for scientific research use only by professionals and is not intended for clinical diagnosis or treatment, food or drug use, or storage in a residential area. For your safety and health, wear lab coats and disposable gloves when handling.