Dihydrorhodamine 123 probe (Intracellular ROS assay) Principle
Some stress conditions like exposure to oxidants or drugs can induce Reactive Oxygen Species (ROS). As a result, ROS induce damage in DNA, protein and lipids, with important effects in cells, leading to oxidative stress.
Cell permeant reagent Dihydrorhodamine 123 (DHR 123) is an uncharged and nonfluorescent reactive oxygen species (ROS) indicator that can passively diffuse across membranes. Oxidation to cationic rhodamine 123 occurs inside cell, which specifically localizes in the mitochondria and exhibits green fluorescence.
It seems that neither the superoxide, the NO, nor the hydrogen peroxide by themselves, are capable of oxidizing DHR 123. According to literature, these ROS, combine with other cellular components such as cytochrome c oxidase or Fe2+ in order to oxidize DHR 123 to its fluorescent derivative Rhodamine 123.
This probe is useful for the detection of reactive oxygen species (ROS) such as peroxide and peroxynitrite. Then, after cell uptake, ROS oxidize DHR 123 into a fluorescent compound.
BQCkit Dihydrorodamine 123 probe presents a solution 1000 times concentrated of DHR 123 ready to use. The protocol for this probe is very simple and contains a stain step, an incubation step and a “semiwash” step. The user can monitor Formation of Rhodamine 123 by fluorescence spectroscopy using excitation and emission wavelengths of 500 and 536 nm, respectively, or by absorbance spectroscopy at 500 nm.
There are three different formats according to the customer needs. These formats are: 250, 500 and 1000 tests.