Home
Resources
Publications
Hydroethidine- And MitoSOX-derived Red Fluorescence Is Not a Reliable Indicator of Intracellular Superoxide Formation: Another Inconvenient Truth

Hydroethidine- And MitoSOX-derived Red Fluorescence Is Not a Reliable Indicator of Intracellular Superoxide Formation: Another Inconvenient Truth

Jacek Zielonka, B Kalyanaraman

Free Radic Biol Med. 2010 Apr 15;48(8):983-1001.

PMID: 20116425

Abstract:

Hydroethidine (HE; or dihydroethidium) is the most popular fluorogenic probe used for detecting intracellular superoxide radical anion. The reaction between superoxide and HE generates a highly specific red fluorescent product, 2-hydroxyethidium (2-OH-E(+)). In biological systems, another red fluorescent product, ethidium, is also formed, usually at a much higher concentration than 2-OH-E(+). In this article, we review the methods to selectively detect the superoxide-specific product (2-OH-E(+)) and the factors affecting its levels in cellular and biological systems. The most important conclusion of this review is that it is nearly impossible to assess the intracellular levels of the superoxide-specific product, 2-OH-E(+), using confocal microscopy or other fluorescence-based microscopic assays and that it is essential to measure by HPLC the intracellular HE and other oxidation products of HE, in addition to 2-OH-E(+), to fully understand the origin of red fluorescence. The chemical reactivity of mitochondria-targeted hydroethidine (Mito-HE, MitoSOX red) with superoxide is similar to the reactivity of HE with superoxide, and therefore, all of the limitations attributed to the HE assay are applicable to Mito-HE (or MitoSOX) as well.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP104821252	Dihydroethidium		104821-25-2	Price

As a specialist in analytical chemical Products, Alfa Chemistry offers consumers a wealth of raw materials and a full range of technologies and services.

QUICK LINKS

HEADQUARTERS

Tel:

Fax:

Email: