GST Antibodies

ijiwaru at wheel.dcn.davis.ca.us ijiwaru at wheel.dcn.davis.ca.us
Wed Oct 23 14:52:07 EST 1996


In article
<Pine.OSF.3.90.961024110536.29247A-100000 at central.murdoch.edu.au>,
eburkala at CENTRAL.MURDOCH.EDU.AU (Evan Burkala) wrote:

> Hi,
>     I was just wondering if any of the many knowledgeable people out 
> there have any idea about the presence of naturally occurring antibodies 
> to the GST fusion protein coded for in the pGEX vector (Pharmacia)? I 
> know it comes from S. japonicum but if one is to use a fusion protein for 
> detection of antibodies to the fusion partner (protein of interest) 
> what is the likelihood of GST antibodies giving false-positives in ELISA 
> in many different animals and even humans. Some parasite experts may 
> comment on the presence of antibodies to GST in natural Schistosoma 
> infection.
> 
> Any correspondence would be appreciated,
> Evan Burkala
> Murdoch University
> Western Australia

Evan,

I do know the naive rabbit sera was free of GST reactive antibodies by
Western blot against the fusion protein and GST from 'empty' pGEX.  Once
immunized, it didn't take long (2 or 3 immunizations) before anti-GST
antibodies were present at a reasonably high titre.  I did Western blots
and immunoprecipitations on material from HeLa cells expressing my protein
of interest and don't remember seeing anything of S.japonicum GST size by
either assay that was significant.  I would guess that at least from HeLa
cells you might expect a low background, but that all depends on the
sensitivity of your ELISA.

Good luck,

Lyle Najita
Plant Pathology
University of California - Davis
>



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