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  • Title: 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.
    Author: Kuroda N, Murasaki N, Wada M, Nakashima K.
    Journal: Luminescence; 2001; 16(2):167-72. PubMed ID: 11312543.
    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.
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