Hello All,
I would like to remove only just ECL and leave most of primary and secondary antibody on the membrane, then store it in -20C for later re-developing the signal with new ECL, may be in days, weeks, or month.
Can it be done?
If yes, what is the method should I use to remove just ECL from membrane? Is it washing? What to use TBS or TBST or Distilled Water or others? and How long of the washing is sufficient?
Thank you very much
Cheerio,
Hi there,
You can freeze the membrane (after washes in TBST) but you will need to probe it again with fresh antibodies after thawing.
The ECL reaction will eventually "burn out". All the substrate will be used up, and no more photons of light will be generated. ECL can only be detected while the chemical reaction is producing photons, so no reaction = no signals.
However, the HRP conjugate has a limited lifetime and the enzyme loses its activity. Although the concentrated conjugate is pretty stable, it is much less stable in dilute solution. I would expect freeze-thaw to destroy the HRP activity on the membrane.
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