Issue 37, 2016

Direct oxidative pathway from amplex red to resorufin revealed by in situ confocal imaging

Abstract

Amplex Red (AR) is a very useful chemical probe that is employed in biochemical assays. In these assays, the non-fluorescent AR is converted to resorufin (RS), which strongly absorbs in the visible region (λabs = 572 nm) and yields strong fluorescence (λfluo = 583 nm). Even if AR is commonly used to report on enzymatic oxidase activities, an increasing number of possible interferences have been reported, thus lowering the accuracy of the so-called AR assay. As a redox-based reaction, we propose here to directly promote the conversion of AR to RS by means of electrochemistry. The process was first assessed by classic electrochemical and spectroelectrochemical investigations. In addition, we imaged the electrochemical conversion of AR to RS at the electrode surface by in situ confocal microscopy. The coupling of methodologies allowed to demonstrate that RS is directly formed from AR by an oxidation step, unlike what was previously reported. This gives a new insight in the deciphering of AR assays' mechanism and about their observed discrepancy.

Graphical abstract: Direct oxidative pathway from amplex red to resorufin revealed by in situ confocal imaging

Article information

Article type
Paper
Submitted
24 Jun 2016
Accepted
22 Jul 2016
First published
29 Jul 2016

Phys. Chem. Chem. Phys., 2016,18, 25817-25822

Direct oxidative pathway from amplex red to resorufin revealed by in situ confocal imaging

P. Lefrançois, V. S. R. Vajrala, I. B. Arredondo, B. Goudeau, T. Doneux, L. Bouffier and S. Arbault, Phys. Chem. Chem. Phys., 2016, 18, 25817 DOI: 10.1039/C6CP04438G

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements