Westrn blot and quantification of protein.

Joan Shields joan at med.unc.edu
Wed Jan 4 10:54:36 EST 1995

Mary asks:
We are currently using western blots to determine expression of a protein.
Our antibody has not been impressive so far.  It has been suggested that
we use immunoprecipitation instead of the western technique.  Can
immunoprecipitation be used to compare amount of protein expression or is
the western the better way to go?  Thanks for your input.   Mary


Immunoprecipatation alone isn't going to give you much, you still need
some way to measure (or at least see) your protein expression.  You could
use immunoprecipitation and then run the sample on a gel and do a western.
It could be that your antibody is reacting with something else along with
what you're looking for.  Immunoprecipitation might get rid of what may be
interfering with the binding you want.  The problem with doing the 
immunoprecipitation before the western is that you may loose some of what
you're looking for (as will happen in any purification step which is
pretty much what an immunoprecipitation step does).  

You could give it a try, probably wouldn't hurt.  You might also want to
look at your technique and solutions as the problem could be there.

Good luck,


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