In article <1jvoehINNql at huon.itd.adelaide.edu.au>, rbonfigl at waite.adelaide.edu.au (Rod Bonfiglioli) writes:
> is there any one out there doing in situ hybridisation work at the TEM level?
> i am doing this on some plant viruses at the moment, and i am hoping to discuss
> some points with other people who are doing similar things. i am attempting to
> locate some viral proteins at the ultrastructural level. anyone out there also
> doing anything like this? i would be pleased to hear from you,
>> thanks in advance for any responses
> Rod Bonfiglioli, Waite Agricultural Research Institute,
> University of Adelaide, South Australia.
> e-mail rbonfigl at waite.adelaide.edu.au
We have done the kind of ultrastructural localization of viral proteins such as
you describe using specific monoclonal antibodies linked to colloidal gold
particles. Antibody binding to the target protein provides specificity for the
target protein; bound antibodies are visible in the EM due to the
electron-dense gold tag. We have used this technique to visualize the locations
of several paramyxovirus proteins in/on the nucleocapsid.
Are there antibodies available for the proteins you plan to examine? And can
the structures you're interested in be made permeable/accessible to antibodies
without disrupting them too much? If this technique sounds useful to you I can
provide references on the methodology.
Kevin W. Ryan
Department of Virology & Molecular Biology
St. Jude Children's Research Hospital
phone (901) 522-0411, fax (901) 523-2622
Internet: ryan at mbcf.stjude.org