HELP. REPROBING BLOTS WITH ECL kit

Theano Markoulides theano at gecko.biol.wits.ac.za
Sat Feb 11 09:20:56 EST 1995


Hi everyone

Does anyone know of the best method of reprobing blots using the ECL kit ?
I need at least five results from each blot, However, I can usually only get 
one or two. Amersham claims that blots can be reprobed with the same probe 
without stripping the blot of the old one. How can this be ? This has never 
worked for me. Can the conventional boiling in 0.1 % SDS method be used to 
strip blots, and then reprobe with the ECL kit ? Amersham also claims that 
the blot should be soaked in the detection reagent overnight to get rid of 
the background from the previous reaction. This also doesn't always work.
Can anyone offer tips or other advice in this regard (other than repeating 
the blot from scratch ?)

Thanks in advance.

Fay Randall



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