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      A cost effective non-commercial ECL-solution for Western blot detections yielding strong signals and low background.

      1 ,
      Journal of immunological methods
      Elsevier BV

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          Abstract

          We compared several alternative ECL solutions for Western blot detection of endogenous proteins in whole cell lysates using inexpensive, commercially available reagents. Starting from an existing protocol based on p-coumaric acid (pCA) as enhancer, we found that the ECL solution containing 4-iodophenylboronic acid (4IPBA) generated strong specific signals and low background chemiluminescence. We optimised the luminol, 4IPBA and hydrogenperoxide concentrations of this 4IPBA-ECL solution. The optimised 4IPBA-ECL solution (100 mM Tris/HCl pH 8.8, 1.25 mM luminol, 2 mM 4IPBA, 5.3 mM hydrogenperoxide) shows a greatly increased signal intensity compared to the initial pCA-ECL protocol and to some commercially available ECL solutions. In addition, the optimised 4IPBA-ECL solution also generates much lower background chemiluminescence than other non-commercial ECL solutions using p-coumaric acid or 4-iodophenol as enhancers. The 4IPBA-ECL solution was stable when stored but had the lowest background when prepared freshly from stock solutions. Thus, we present an optimised protocol for a well-performing inexpensive ECL solution which is an alternative to expensive commercial ECL solutions and which achieves a better signal and lower background than the commercial solutions tested.

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          Author and article information

          Journal
          J Immunol Methods
          Journal of immunological methods
          Elsevier BV
          0022-1759
          0022-1759
          Jan 10 2007
          : 318
          : 1-2
          Affiliations
          [1 ] Laboratoire de Biologie et Physiologie Intégrée, Fac. Science Techno. and Com., Université du Luxembourg, 162a, Avenue de la Faïencerie, L-1511 Luxembourg, Luxembourg. claude.haan@uni.lu
          Article
          S0022-1759(06)00294-8
          10.1016/j.jim.2006.07.027
          17141265
          b11c67d9-826c-4f28-91b4-543695898b3c
          History

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