Immunoprecipitation-quantitative??

Martin Offterdinger a8803349 at unet.univie.ac.at
Tue Jan 7 02:25:24 EST 1997


On Fri, 03 Jan 97 17:20:38 GMT,  (Dima Klenchin) wrote:

>In article <32ccbd2b.481197 at news.univie.ac.at>,
>   a8803349 at unet.univie.ac.at (Martin Offterdinger) wrote:
>->Hi
>->I am wondering if it is possibble to set up a quantitative
>->immunoprecipitaion to analyse the regulation of expression of a
>->particular cell surface receptor: What I would like to do is to
>->immunoprecipitate the receptor and do a western afterwards with
>->hormone treated and untreated cells and compare the relative
>->expression. Do you think that this would be an appribiate approach??
>
>I'd be very sceptical of such approach and certainly would not trust results
>quantitatively. One poorly reproducible and highly non-linear technique
>(IP) on top of another highly non-linear (western). Most people have enough
>troubles introducing all sort of errors in "quantitative" westerns. 
>However, if nothing else is feasible, I'd go for western alone, omitting IP
>as essentially redundant step. 
>
>- Dima
>
The problem is that Western alone would not be sufficient sensitive to
detect the receptor-therefore I do not think that IP is a redundant
step-I would like to use it essenitally to increase sensitivity.
Martin



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