IUBio

Assay protein bound to ELISA

M. Doherty M_Doherty at NIH.gov
Tue Sep 12 10:04:53 EST 1995


In article <432or5$eu1$1 at mhafn.production.compuserve.com>, Geoffrey D.
Wheelock <74710.2416 at CompuServe.COM> wrote:

> Someone asked how to assay protein bound to immulon 2 ELISA plates. 
> My reader trashed the original article so I am replying from memory.
>  The amount of protein bound to the plate will be very small, circa 
> 100 ng or less, so you need a really sensitive assay.  I have done 
> it by iodinating the protein or peptide, determining the specific 
> activity of the protein, binding to the plate, washing, and counting
> the wells in a gamma counter.  Of course, if you are not using strip
> wells, breaking up the plate into individual wells is problematic :)
>  Hope this helps.  Geoffrey D. Wheelock

I took the same route.  Given the small amounts of protein bound,
iodination seemed to be the most viable technique.  However, lacking
plates with individual wells, I simply cut the plate into bits with a bone
saw and dropped each well into a regular vial for assay.  Messy and
tedious, but effective!

Cheers, Mark Doherty



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