western-quantitation??

Tobias Straub tobias.straub at mail.uni-wuerzburg.de
Wed Aug 26 13:41:16 EST 1998


In article <35e40dba.2700387 at news.univie.ac.at> ,
a8803349.nospam at unet.univie.ac.at (Martin Offterdinger) wrote:

>I would like to know if you would consider it as science to quantitate
>bands obtained by enzymatic Western Blotting with a densitometer. I
>would like to study the expression of a certain protein under the
>influence of hormones and compare it to control treated cell extracts.
>I know if you put the film under the densitometer you will get numbers
>(per cent of control) anyway. But is it good practice to conclude from
>the relative band intensity in such an experiment to the relative
>protein expression or is it just numbers that do not mean anything?
>I am looking forward to discussion.

I think another major problem of the western blot is that you cannot be
shure that all your protein actually transferred and bound to the membrane.
Furthermore I'm not quite shure if the transfer efficiency is all the same
within one gel. In my opinion the western blot maybe used for
semiquantitative analyses but never more than that.
Tobias
------------------------------------------------
Tobias Straub
Medizinische Poliklinik, University of Wuerzburg
Klinikstrasse 6-8
97072 Wuerzburg
Germany
Phone: +49-931-2017085
Tobias.Straub at mail.uni-wuerzburg.de



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