TITLE

Amperometric detection in TMB/HRP-based assays

AUTHOR(S)
Fanjul-Bolado, Pablo; González-García, María Begoña; Costa-García, Agustín
PUB. DATE
May 2005
SOURCE
Analytical & Bioanalytical Chemistry;May2005, Vol. 382 Issue 2, p297
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
3,3′,5,5′-Tetramethylbenzidine (TMB) is the most commonly used chromogen for horseradish peroxidase (HRP) and so its performance as an electrochemical substrate was evaluated. Measurements of HRP activity in solution were carried out by using an amperometric detector coupled to a flow injection analysis (FIA) system. The enzymatic product was easily detected at a potential of +0.1 V (vs. Ag-pseudoreference electrode) at a bare screen-printed electrode placed in a homemade electrochemical flow cell. A high flow rate (4.3 mL min−1) of 0.5 M H2SO4 was used to obtain repeatable signals and a short analysis time. The detection limit achieved after 15 min of incubation was 2×10−14 M of HRP. The applicability of the amperometric detector to ELISAs was demonstrated by using a commercially available kit for the quantification of interleukin-6 (IL-6) without modifying the kit manufacturer’s protocol or the reagents for this test.
ACCESSION #
17252632

 

Related Articles

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics