ECL Western troubles

Meirion Jones m.l.jones at uk.ac.birmingham
Wed Sep 18 11:19:56 EST 1996


> Jose Bonner <jbonner at bio.indiana.edu> wrote:
> >
> > We're having trouble with Western blots probed by ECL.  Films sometimes 
> > come up blank, even though antibodies and reagents are OK.  Sometimes a 
> > 5-minute exposure is fine, but a 1-hour exposure (started just after) is 
> > blank--not even any background.  What can interfere with ECL?  The 
> > problem comes and goes; good blots interspersed with blanks.  Reprobe 
> > the blanks, and sometimes they come up good; reprobe the good ones, 
> > sometimes they come up blank.  Anyone else have this problem?

I'm having the same problems with my ECL Westerns, more often being blank,
or with very poor signals, than working. The developer is working OK and
the only reason we could come up with was that something must be
interfering with the reagants in some way. The reagants are made up in
sterile containers though so we are a bit stumped as to what the problem
could be. 

I may have missed the start of this thread so I would appreciate any
thoughts anyone has on the problem.

Thanks in advance

Meirion Jones
Biological Sciences
Birmingham University



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