homemade receipt for ECL reagents

[OKADA Yasushi]

In article <34t9js$s0f at network.ucsd.edu> T. S. Pillay <tpillay at ucsd.edu> writes:
> In article <34ndav$pj0 at msuinfo.cl.msu.edu> Your Name Here,
> you at somehost.somedomain writes:
> >
> >Hi netters, 
> >Does anybody have information about how to prepare the reagents for
> enhanced 
> >chemiluminescence (ECL) reactions?	
> >Any help will be greatly appreciated. Thanks in advance.
> 
> Start off with luminol- available from Sigma (needs to be dissolved in
> DMSO-I think). Then you need an enhancer-  for further information see
> papers and reviews by Kricka and others.

Here is the recipe I use,

Stock solns.
a) Luminol stock: Luminol (Sigma) 4mg/ml in DMSO.
b) Enhancer stock: p-iodophenol (Aldrich) 1mg/ml DMSO.
c) Tris Buffer: 100mM Tris.HCl pH7.5
d) H2O2: 3% H2O2

Stock solns a) and b) should be kept in -20deg for <6 months.

Working soln. (Prepare just before use)
a) 0.5ml, b) 0.5ml, c) 2.5ml, d) 25ul, ddw 1.5ml (final 5ml)

This is very cheap, and as sensitive as ECL kit.

Reference: Schneppenheim et al (1991) Electrophoresis 12:367-372.

Hope this helps.

--
Yasushi Okada, MD.
($@2,ED9/;V(J <= Japanese Kanji)
Department of Anatomy & Cell Biology
University of Tokyo
($@El5~Bg0e!&Bh#12rK6(J <= Japanese Kanji)
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