ECL reagents

Vincent Solomon vsolomon at fas.harvard.edu
Sat Nov 2 12:12:35 EST 1996


Jeff \"Newt\" Sekelsky (fznewt at peseta.ucdavis.edu) wrote:
: 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

: ----------------------------------------------------------------
: On 1 Nov 1996, A. Piccin wrote:

: > Hi everybody.
: >=20
: >        Like many others nowadays, I am using the Amersham ECL kit for=20
: > immunodetection on Western blots. And I am bit upset by its price. Amersh=
: am
: > (clever people) reckons that one should use 0.125 ml/cm 2 of membrane. Th=
: at works
: > out to about 8-10 blots in my case, paying =A398 for the kit. Actually I =
: started
: > to use much less than the recommended volume of reagents, and spreading i=
: t 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 solu=
: tions.
: > Does anybody know the recipes for a reliable luminescent detection reagen=
: t?
: >=20
: > Thanks for your help
: > Alberto

--
	
I use the recipe given above.   It works well.

Vince



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