Since ECL is provided from many companies viz Thermo, biorad etc. which is bit costly. Can anyone provide with me with the working laboratory protocol for ECL preparation for Western Blotting?
Thanks in advance
Totally agree with Raquel, the link tells you how to do it. We tend to make the solution fresh each time because it takes only a few minutes when you have the luminol and coumaric acid solutions aliquoted in the freezer, hydrogen peroxide in the fridge and 1M Tris-HCL pH 8.5 on your bench. If you do westerns several times a week, please notice that every time you open and close the hydrogen peroxide bottle you will get progressive reduction of the peroxide concentration (humidity gets in, peroxide gets out). We tend to replace the hydrogen peroxide every 12 months even if the bottle is still quite full, and we gradually increase the amount of peroxide (like 10% every three months) until we buy a fresh bottle (its cheap).
Hello Jürgen Denecke,
I am attaching the result file for home made recipe of ECL which you suggested. I observed a bit of background.
How to reduce the background ?
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