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090			_aB-12529
245	1	0	_aA cost effective non-commercial ECL-solution for Western blot detections yielding strong signals and low background
490	0		_vJournal of Immunological Methods, 318(1-2), p.11-19, 2007
520	3		_aWe 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.
650	1	4	_aENHANCED CHEMILUMINESCENCE
650	1	4	_aWESTERN BLOT
650	1	4	_a4-IODOPHENYLBORONIC ACID
650	1	4	_aP-COUMARIC ACID
650	1	4	_a4-IODOPHENOL
700	1	2	_aHaan, C.
700	1	2	_aBehrmann, I.
856	4	0	_uhttps://drive.google.com/file/d/1ofQoNnWiZsIk0xw6hyqoSDPrlVy1RKDm/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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