Hi Mohsen

Although you can run Native PAGE instead of SDS-PAGE to exclude the effect of SDS, I doubt extracting from the PAGE is a good strategy because you also need to eliminate A/BA from your sample.

I'm pretty sure that It will be better to purify your sample by chromatography.

If you are looking for a specific protein, you can also purify it by Co-IP and the final elution contains your desired protein in pure form.

The classical procedure is to run your sample on SDS-PAGE, fix, stain and destain the gel and then cut out the band of interest. Wash with 2 changes of PBS, mash it up thoroughly in adjuvants (complete first time, incomplete for boosters) and then inject. Contrary to acrylamide, polyacrylamide is not toxic, but serves as a depot for the antigen. If you are very short of antigen, you can inject directly into a lymph node at the hind leg of the rabbit, this is a surgical procedure, so you'll need a vet to carry it out.

For all antibody-related procedures, check out ISBN 9780879693145, which is still the authoritative reference. And of course, you need permission from your local animal use committee for any such experiments.

Masood Sepehrimanesh

Thank you for your comment, Masood. I'm aware of other purification methods, but I have concluded this suits me the best.