On 28 Dec 1995 22:00:19 GMT,
Rob Chen <rchen at orion.oac.uci.edu > wrote:
>>Does anyone out there have experience with chicken IgY antibodies? I
>understand that there are many advantages, such as short immunization schedule
>and very high yield for typical proteins. What I would like to know is how
>does IgY compare to mammalian IgG when used for immunocytochemistry,
>precipitation, western blotting, etc. Are they comparable?
It is somewhat difficult to get the IgY completely cleaned up (all the
lipid removed) but when this is done they work fine on westerns. They
also work well in IPs. I did not get any experience with
immunocytochemistry. The major lack of comparability is that IgY is
not recognized by proteins A or G and most of the secondary Abs that
are commercially available are raised to serum IgG which only mildly
cross-reacts with IgY. It was best to make my own secondary antibody,
but then this results in labeling it myself or using a triple probing
system (IgY, rabbit anti-IgY, then protein A). This can be a
problem resulting in a low signal in IPs.
>How about the cost
>of immunization, purification, and secondary antibodies?
The cost of housing here was comparable to rabbits, though I had to do
the immunization myself (whereas a service will do it with the
rabbits). Howver, no bleeding is required, since one only collects the
eggs. Purification costs are fairling low.
In the end I abandoned chickens due to the inconvenience (no
immunization sevice, and the farm is about 8 miles away) and the lack
of ease of use due to the need for secondary antibodies.
Purdue University, West Lafayette, IN
ashendel at aclcb.purdue.edu