ECL Western blotting detection kit - (Dec/08/2004 )

Hi,

Has anyone tried using this kit from Amersham (ECL Western bolotting detection reagent RPN2209)? Can the image on the membrane be captured using light box instead of on X-ray films?

Thanks.

If by light box you mean something like the Epichemi II darkroom (UVP Lab Products, UK) which has a CCD camera with appropriate filters or Biorads VersaDoc system then the answer is yes. I actually found the Epichem more sensitive that film

Thanks DevGrp. Unfortunately my lab doesn't have any of those.

Well in therory any sensitive CCD camera with an appropriate filter and in some kind of dark room / dark box should work.
The only problem would be getting software to allow you to collect the image over several minutes (5-15min)

As an aside, if you want to save your lab lots of money, then you can make your own ECL reagents.

they work just as well as ECL reagents but at a fraction of the cost.

Solution 1

1ml 250mM luminol
0.44ml 90mM p-coumaric acid
10ml 1M tris pH 8.5
up to 100ml with milliQ water

Solution 2
64ul 30% hydrogen peroxide
10ml 1M tris pH8.5
up to 100ml with milliQ water

you use an equal volume of each solution on a blot, typically 1ml of each and leave on for 1 minute.
As the above solutions don't have preservatives, they won't last for months and months but I've used them up to 2months after preparing and they still worked fine