Best ECL detection reagents - (Oct/03/2012 )
What type of ECL detection reagents are you using and why? There are so many of them today it's difficult to choose. I'm looking for high sensitivity and long-lasting signal.
Thanks in advance!
In my experience the ECL you is very dependent on your on you protein of interest, I work on proteins which are expressed at very low levels under native conditions so we use this http://www.piercenet.com/browse.cfm?fldID=01041102 because it allows us to do long exposures and is very sensitive. However if you use it on proteins which are expressed at moderate to high levels it will give you very strong bands unless you do very short exposures
This may draw great deal of derision but for westerns of well expressed proteins I use a Santa Cruz ECL (http://www.scbt.com/datasheet-2048-western-blotting-luminol-reagent.html) I have no particular reasons expect that it has always worked for me and is what I have always used and it is cheap!
But recently we have started to use quantum dot based reveals with a great deal of success due to the strength of the signal we can load less protein and so produce much cleaner blots than we ever previously were able to obtain and multiplexing is possible. However this does require addition pieces of equipment but in its most basic form can be done on a transilluminator.
Hope this helps
We use this for proteins with very low expression http://www.piercenet.com/browse.cfm?fldID=01041106 and this for moderately expressed proteins http://www.piercenet.com/browse.cfm?fldID=560371F1-8179-4323-92AD-4A96296F8929