Detection of immunoglobulin E using an aptamer based dot-blot assayReport as inadecuate




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Chinese Science Bulletin

, Volume 58, Issue 24, pp 2938–2943

First Online: 24 April 2013Received: 15 September 2012Accepted: 23 November 2012DOI: 10.1007-s11434-013-5702-9

Cite this article as: Wang, Y., Ye, Z. & Ying, Y. Chin. Sci. Bull. 2013 58: 2938. doi:10.1007-s11434-013-5702-9

Abstract

A novel aptamer based dot-blot assay for the detection of immunoglobulin E IgE was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immobilized onto a nitrocellulose membrane substrate. Avidin conjugated horseradish peroxidase was introduced onto the membrane through the biotin-avidin system to catalyze the hydrogen peroxide mediated oxidation of 3,3′,5,5′-tetramethylbenzidine, thereby producing the blue-colored insoluble product. The intensity of the dots increased as the concentration of IgE increased. The spot intensity was quantified using a simple portable instrument. A linear response relationship between the spot intensity and the concentration of IgE over the range of 50 nmol-L to 1 μmol-L was obtained. The detection limit for IgE using the aptamer-based assay was 2.89 nmol-L. This assay was found to discriminate IgE from non-target proteins such as thrombin, bovine serum albumin and immunoglobulin G.

Keywordsimmunoglobulin E dot-blot assay aptamer biotin-avidin system This article is published with open access at Springerlink.com

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Author: YiXian Wang - ZunZhong Ye - YiBin Ying

Source: https://link.springer.com/







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