Good rabbits or bad rabbits
M.Mason at botany.uq.edu.au
Sun Feb 16 02:45:55 EST 1997
To who ever,
I am a PhD student at the University of Queensland, and my project involves
raising antibodies against a particular protein.
I started off injecting two rabbits with Freunds adjuvant mixed
with 100ug of un-dialysed protein which contained approximately 1M Urea. After
4 sets of injections (conducted every 2 weeks) the blood serum was not
detecting any proteins on a Western Blot at 1/100 dilution. After this I
decided to dialyse the protein against PBS, and repeat the immunization
protocol again, using 200ug of protein on the same two rabbits. Other than the
controls, and the purified antigen, no bands were seen on the Western. I
continued to inject every two weeks for another 6 weeks. After this I started
to get some faint bands on a Western using 1/100 diluted serum.
I do know however that the protein I am trying to detect is on the western
membrane. So it can not be a simple fact of it not coming out in the
Could it be that I have bad rabbits?, a little to old maybe? or did the Urea
in the protein solution destroy antibody production? or could it be the
I would greatly appreciate it if anyone can come up with some sugguestions, as
I am now starting to inject two new rabbits with dialysed protein.
Thanks in advance
M.Mason at Botany.uq.edu.au
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