Application of an enhanced luminol chemiluminescence reaction using 4-[4,5-di(2-pyridyl)-1H-imidazol-2-yl]phenylboronic acid to photographic detection of horseradish peroxidase on a membrane

Luminescence : the Journal of Biological and Chemical Luminescence
N KurodaKenichiro Nakashima

Abstract

Photographic detection of horseradish peroxidase (HRP) on a membrane by the luminol-hydrogen peroxide-HRP chemiluminescence reaction using 4-[4,5-di(2-pyridyl)-1H-imidazol-2-yl]phenylboronic acid (DPPA) as an enhancer is described. The method is based on the long-lived chemiluminescence emission obtained by using DPPA. Under the optimum conditions, as little as 0.10 ng (ca. 2.3 fmol) and 0.20 ng (ca. 4.6 fmol) per spot of HRP on a membrane were detected as visible spots with exposure time of 60 and 10 min, respectively, by using an instant photographic film and a camera luminometer. The proposed method was highly sensitive and was successfully applied to the detection of HRP conjugates as an alternative to the colorimetric method using a chromogenic substrate in a commercially available assay kit of Western blotting.

References

Jan 1, 1986·Methods in Enzymology·G H Thorpe, L J Kricka
Sep 1, 1968·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·P K Nakane
Oct 1, 1982·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·S M Hsu, E Soban
Dec 22, 1999·Luminescence : the Journal of Biological and Chemical Luminescence·N KurodaK Nakashima
Feb 1, 1965·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·R C GRAHAMM J KARNOVSKY

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Citations

May 1, 2013·Luminescence : the Journal of Biological and Chemical Luminescence·T IchibangaseN Kuroda
Aug 30, 2002·Analytical Chemistry·Rezik A AgbariaIsiah M Warner

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