ECL reagents

Jeff "Newt" Sekelsky fznewt at peseta.ucdavis.edu
Fri Nov 1 20:52:52 EST 1996


I've not personally tried this yet, but a friend sent this recipe:

Stock solns.- store at 4, they will freeze so thaw at 37.
1. 250mM Luminol (Sigma A-8511, 5g)
        0.44g/10mL DMSO (keep light tight)
2. 90mM p-Coumaric acid (Sigma C-9008, 10g--there may be smaller sizes)
        0.15g/10mL DMSO

Solution A: (keep at 4, light tight)
1mL Luminol stock
0.44mL p-Coumaric acid stock
10mL 1M Tris-HCl, pH 8.5
bring 10 100mL

Solution B: (keep at 4, but it goes off quickly so make weekly)
10mL 1M Tris-HCl, pH 8.5
61uL 30% H2O2
bring to 100mL

Mix Solutions A and B 1:1 just before use

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On 1 Nov 1996, A. Piccin wrote:

> Hi everybody.
> 
>        Like many others nowadays, I am using the Amersham ECL kit for 
> immunodetection on Western blots. And I am bit upset by its price. Amersham
> (clever people) reckons that one should use 0.125 ml/cm 2 of membrane. That works
> out to about 8-10 blots in my case, paying £98 for the kit. Actually I started
> to use much less than the recommended volume of reagents, and spreading it on
> the membrane with a sheet of Saran wrap. About 2 ml of reagent is usually OK
> for a 130 cm2 blot. But I would be happier if I could make up my own solutions.
> Does anybody know the recipes for a reliable luminescent detection reagent?
> 
> Thanks for your help
> Alberto
> 
> 
> 




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