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

 -  Journal of Immunological Methods, 318(1-2), p.11-19, 2007 .
<br/><br/> 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 noncommercial ECL solutions using p-coumaric acid or 4-iodophenol as enhancers. The 4IPBA-ECL solution was table when stored but had the lowest background when prepared freshly from stock solutions. Thus, we present an optimised protocol for a wellperforming 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. 
<br/><br/><label>Subjects--Topical Terms: </label><br/>ENHANCED CHEMILUMINESCENCE<br/>WESTERN BLOT<br/>4-IODOPHENYLBORONIC ACID<br/>P-COUMARIC ACID<br/>4-IODOPHENOL