In article <1383228364-67665670 at post.miyazaki-med.ac.jp>,
Seung Kyu Park <eisei at MACQM.MIYAZAKI-MED.AC.JP> wrote:
>Dear Everyone:
> I have interest in making antibodies against highly conserved
proteins among the species. Because of the conservation, other methodology
will be needed for making antibody. But i have no experimental ideas about
it. Any kinds of informations or directions would be appreciated.
(the above was edited to shorten the line length)
If you've already had problems and it doesn't matter if your antibody is
polyclonal, you will have better luck by raising polyclonal serum antibody
in chickens. If you need to contact someone who does this once in a while,
contact me by email and I'll hunt down her phone number. She's had good luck
with other proteins that were too highly conserved to raise antibodies
to in other mammals. Obviously, the greater evolutionary distance between
birds and mammals helps A LOT. You can purchase secondary antibodies to
the chicken antibodies from Southern Biotech.
There is, in fact, a chicken fusion partner available from researchers in
Japan. If you're interested then check Medline because I don't have the
paper anymore. There are a few very serious problems with it (which we
were warned of by the authors). The first, less serious problem is that
the line does make its own heavy or light chain (I forgot which) and
the second is that the hybridomas you generate will be VERY unstable,
requiring a lot of _constant_ subcloning. These warnings came true for
my friend, who could deal with with former problem but I thought she'd
kill herself over the latter (she's really good at what she does and
had PLENTY of unstable hybridomas to deal with - I hope she got some
medication for the nervous condition it gave her :)
Personally, I'd stick to the chicken polyclonal antisera and skip the
fusions.
Stacy
