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[Protein-analysis] Re: affinity columns

Allison via proteins%40net.bio.net (by allison from nospam.com)
Wed Mar 28 15:27:46 EST 2007

Dan Guire wrote:

> Hi Barbara,
> The main point has already been made:  Your anti rabbit alphatase conjugate
> can only detect the rabbit antibody down on the Maxisorp plate.  It doesn't
> matter whether there is antigen for the capture antibody or not.  No doubt a
> negative control, leaving out only the mouse lysate, would show you that. 
> The series Allison outlined looks fine; the enzyme-conjugated anti mouse
> shouldn't bind unless there is mouse antibody bound to antigen there.  My
> point is that if you already have biotinylated anti ENAC of one species, you
> can capture any antigen with the non-biotinylated anti ENAC of the other
> species, then detect with the biotin and a streptavidin-enzyme conjugate.
> And a negative control is a must :-)
> DanG
> P.S. Do you have any affinity column experience by any chance?  I could use
> some help in that arena...

I have done a bit....what is your question?  If I can't help I am sure 
someone else can.

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