"T. Andersen" wrote:
>> I have been told some years ago that it is possible to cut out protein bands
> from SDS-PAGE gels, dry the gels and then use them for direct immunization.
> Does anyone have a detailed protocol on this or a protocol on using proteins
> transfered to a membrane for immunization?
> Thanks!
There is not need to dry the gels. I have a protocol here from: Dunbar,
B.S. Two-dimensional electrophoresis and immunological techniques"
Plennum press, NY, 1988. They cut 2-3 coomassie blue stained spots
(10-200ug protein) from the gel, they mince them in small pieces in
buffer or water and then they pase it back and forth between two
syringes connected with a tube, until everything breaks. Then they
emulsify the thing with adjuvant and they inject it to the rabbit.
Actually proteins used directly with the acrylamide matrix are claimed
to be more immunogenic than the electroeluted or purified thing.
Oswaldo
