Diluted ECL Reagent ?
rwange at helix.nih.gov
Mon Mar 14 23:53:48 EST 1994
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.
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.
both Amersham's HRP-sheep anti-mouse
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