Best substrate for Western-HRP

Andreas Simm simm at klin-biochem.uni-wuerzburg.de
Mon Jun 21 02:04:42 EST 1999



"Paul S. Brookes." wrote:

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> "Duolox" from Vectorlabs works for both AP and HRP, and the signal lasts for up
> to 8 hours.  The only problem is that I find its quite sticky on PVDF and so
> the background is high unless you wash it several times in PBS/T20.  ECL Plus
> from Amersham/Pharmacia is less sticky but the signal fades faster.  There are
> also several protocols on the web for home-made ECL reagents using
> Luminol/DMSO/H2O2 for a fraction of the cost.
>
> Regards
> PSB
>
> _________________________________________
> Dr. Paul S. Brookes.            (brookes at uab.edu)
> UAB Department of Pathology,   G004 Volker Hall
> 1670 University Blvd., Birmingham AL 35294 USA
> Tel (001) 205 934 1915     Fax (001) 205 934 1775
> http://peir.path.uab.edu/brookes
>
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> <html><div>&quot;Duolox&quot; from Vectorlabs works for both AP and HRP,
> and the signal lasts for up to 8 hours.&nbsp; The only problem is that I
> find its quite sticky on PVDF and so the background is high unless you
> wash it several times in PBS/T20.&nbsp; ECL Plus from Amersham/Pharmacia
> is less sticky but the signal fades faster.&nbsp; There are also several
> protocols on the web for home-made ECL reagents using Luminol/DMSO/H2O2
> for a fraction of the cost.</div>
>

Just one additional question,
how often do you wash before you add ECL substrate?
Did you really get a blank background by washing your
membran before adding ECL substrate?

Andreas





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