ECL problem

John A. Newitt newitt at ncifcrf.gov
Tue Apr 18 16:27:45 EST 1995


In article <1995Apr18.161247.1 at hkucc.hku.hk>, h8719263 at hkucc.hku.hk wrote:

> The problem is: in 9 out of 10 expts my blots are very very dirty. And the 
> 'dirt' is in the form of well-defined lines or 'scratches' across the filters.
> The line pattern is usually continuous, even I cut the filters into different 
> strips for different antibodies. The problem doesn't seem to be due to
> inadequate blocking and washing (as we did that excessively but it happened),
> nor is it due to physical damage to the nitrocellulose itself (as we have
> handled the filters as gently as possible). 

I have seen similar "dirtiness" and could always attribute it to a primary
antibody.  That is, some primaries do it consistently, while others never
do.  Have you seen the same thing with multiple primary antibodies?  If
it's just a problem with one antibody, you may be able to clean up the
blots by affinity purifying the primary antibody.

John A. Newitt  <newitt at ncifcrf.gov>
National Institutes of Health
Bethesda, Maryland  USA



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