[western blotting] reproducible detection in western blotting

Pow Joshi via methods%40net.bio.net (by pow.joshi At gmail.com)
Tue Nov 21 16:14:12 EST 2006


On 21 Nov 2006 10:42:15 -0800, Lechu <lech_kaczmarczyk At yahoo.com> wrote:
> What you would reccommed as an alternative to ECL systems + standard
> autoradiography film? I start to find  film developing pretty boring
> and time-consuming, and I always have problems with the right
> adjustment of exposure time to get reproducible results. Lifetime of
> luminescence is also pretty short. Any suggestions as for reasonable
> alternatives? Maybe some fluorescence-based methods?

you could use regular "substrate-enzyme" colour developing methods...
(please check up the substrates for alkaline phosphatase and HRP in
"Molecular Cloning" by Sambrook and Maniatis; I don't remember it on
the top of my head ...been a while since I used them.)....they work
well, if you have high specificity antibodies.

Alternatively, you could use the "Licor" machine, if you have access
to it, which uses some special IR tagged secondaries.... I have seen a
demo of it, and have'nt really used it myself to particularly know how
it works.

best,
pow
> Best wishes, Lech
>
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