Diluted ECL Reagent ?DIR

fleming at guvax.acc.georgetown.edu fleming at guvax.acc.georgetown.edu
Fri Mar 18 16:17:04 EST 1994

In article <1994Mar15.045348.10898 at alw.nih.gov>, rwange at helix.nih.gov (Ron Wange) writes:
> gchacko at magnus.acs.ohio-state.edu (George W Chacko) writes:
> : 
> : Has anyone tried to dilute Amersham's ECL reagent? The kind of signal
> : I get usually requires 3-10 secs exposure and it occurred to me that
> : if I could dilute the reagent in TBS-T or something I wouldn't use it
> : up as fast.
> : 
> : George
> : 
>      I have not tried to dilute ECL reagent; however, I have found
>      (despite Amersham's protests to the contrary) that the ECL reagent
>      can be reused if stored at 4 degress between uses.  We routinely 
>      use a single batch of ECL reagent for one week with good results.
>      Should a batch of ECL reagent go bad, one need only re-expose the 
>      blot to fresh ECL reagent.
>      I suspect that your 3-10 sec exposure times are due to your primary
>      or secondary antibodies being to concentrated.  We routinely use
>      Amersham's HRP-sheep-anti-mouse-IgG and HRP-goat anti-rabbit-IgG
>      at 1:15,000 dilution in TBST.
>      D
>      D
>      both Amersham's HRP-sheep anti-mouse

I have for a long time reused the ECL reagents. Ususally a 1:5 
dilution of the combined reagents in water or TE -whatever I grab at 
the time. No increase in bkgd, just longer exposures. I have also done 
the same with other manfacturers reagents -no problems.
good luck\

rob powell

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