GST fused protein as immunogen?

Frank O. Fackelmayer Frank at Fackelmayer.de
Thu Jun 14 08:10:31 EST 2001



Jinguo Wang wrote:
> 
> Anyone has experiences with GST fused protein as immunogen? My
> recombinant GST fused protein is in inclusion body form and the rabbit
> got response against the fused whole protein. However, I have some
> trouble with separating the abs against the GST and my real target
> protein. After absorption with the rGST, it seems I lost everything..

that is most likely because your rabbit produced antibodies only to GST.
In our hands, GST was so highly immunogenic that we also didn´t get any
titer for the other fusion partner (tested with two unrelated proteins).
I guess the same applies to your experiment. Try subcloning into a
different vector, eg. one that makey His-tagged protein. The his Tag is
known to be very weakly immunogenic, and in our hands his-tagged
proteins made good antibodies (tested with >10 proteins)

Frank




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