ECL Western troubles

Lely Chow lelyc at u.washington.edu
Sun Sep 22 16:40:12 EST 1996


I have had the same problems with ECL.  We use the ECL kit for Northern
blot signal detection (use dig-dCTP in random priming).  We have had the
experience where we get a signal, but after stripping and reprobing, we
cannot detect the same signal.  We gave up trying to figure out what was
going on and switched back to P32 probes, but we thought the problem was
related to the used of DIG.

--Lely


On 18 Sep 1996, Meirion Jones wrote:

> > 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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