Western
Ian A. York
iayork at panix.com
Wed Aug 2 09:14:40 EST 2000
In article <39882A4B.ABB17C14 at biocomp.unl.edu>,
Chris LaRosa <clarosa at biocomp.unl.edu> wrote:
>
>I have used BCIP/NBT westerns for years. Can you elaborate on the
>advantages of these chemiluniescence systems. If they are most
>commonly used there must be a reason..... Better quality images??
I switched from the various colourimetric to chemiluminescent five or six
years ago. In general, the chemiluminescent are more sensitive and have
lower background, though the lower background is a relatively recent
improvement. If antibody is limiting, or expensive, you can usually get
away with lower first and second antibody concentrations. There are other
advantages: The record is more permanent, it's easier to photograph and
scan, and it's easily possible to get multiple exposures: if you have
bands of different intensities you can get several from one blot. It's
also simple to run multiple sequential probes on a single blot: I
routinely run three or four antibodies sequentially on each blot.
But if your system ain't broke, don't fix it.
Ian
--
Ian York (iayork at panix.com) <http://www.panix.com/~iayork/>
"-but as he was a York, I am rather inclined to suppose him a
very respectable Man." -Jane Austen, The History of England
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