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