npv at mani.unet.umn.edu
Tue Jan 10 14:30:08 EST 1995
I made antibodies in chickens and have isolated the IgGs from egg
yolks, but I was not entirely pleased with the results. I had high
background reactivity of the pre-immune yolk IgGs from the three
chickens I used, and it was therefore absolutely essential to include
a control reaction with the pre-immune IgGs in all experiments.
However, this problem may depend on the conditions under which the
soon to be immunized chickens are kept, and an affinity isolation of
the specific IgGs should get around the problem.
Also, if you are considering immunoprecipitation with the antibodies,
you should be aware that chicken IgGs do not react with protein A or
protein G, which therefore cannot be used to amplify the
immunoprecipitation (unless you use an anti-chicken IgG antibody from
mammals first, as I did). However, there are now other tools on the
market that may get around this problem. Promega (Madison, Wisc)
offers an immobilized anti-chicken IgY (IgG) for immunoprecipitation,
which may be useful.
nora at molbio.cbs.umn.edu
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