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ECL Western blotting detection kit


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4 replies to this topic

#1 SilverSta

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Posted 08 December 2004 - 01:36 AM

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.

#2 DevGrp

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Posted 08 December 2004 - 08:59 AM

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

#3 SilverSta

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Posted 08 December 2004 - 05:55 PM

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

#4 DevGrp

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Posted 09 December 2004 - 01:46 AM

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)

#5 patirl

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Posted 26 January 2005 - 10:53 AM

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




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