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Product Reference
A cost effective non-commercial ECL-solution for Western blot detections yielding strong signals and low background.
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| Author |
Claude Haan, Iris Behrmann |
| Citation Information |
Journal of immunological methods, 318:11-9 (2007) |
| Keywords |
Blotting, Western, Boron Compounds, Cell Line, Tumor, Coumaric Acids, Humans, Hydrogen Peroxide, Iodobenzenes, Janus Kinase 1, Luminescence, Luminol, Solutions |
| Related Products |
MAB1501 |
| Pub Med ID |
17141265 |
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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.
