Problems with densitometry!....for WESTERN BLOTS

Jeff Rawlings rawlings at
Fri Mar 24 13:34:28 EST 1995

In article <gjmadlam-2203951154280001 at>,
gjmadlam at (Gerard ) wrote:

> I've just made a strange discovery when developing by blots.  I found that
> when I expose my blots to film, after using ECL, the bands that I get for
> shorter exposures aren't always "lighter" than the bands I get for longer
> exposures.  They are sometimes darker!!!!  This also happens in reverse
> (of course).  I was just wondering if anyone else has had this problem
> and, if so, what solutions are there.  Thanks.

I'm not sure if the chemiluminescent systems are linear (they might be - I
just don't know). The solution I used was to use I-125 labeled
secondaries, and develop the blots in a phosphorimager. Both of those
steps should be linear and quantitative.


Jeff Rawlings                                   Internet Consultant
rawlings at                                             and
rawlings at                         Research Biologist

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