Re: double IHC/in situ - a refinement.
|From:||Marek Malecki <email@example.com>|
Gayle et al.
Sorry. Below, there is a refinement.
Childs, GV. Simultaneous identification of a specific gene protein product
and transcript using combined immunocytochemistry and In Situ Hybridization
with non-radioactive probes. In Science of Specimen Preparation for
Microscopy and Microanalysis . Malecki and Roomans (eds.) by Scanning
Microscopy International vol. 10, 17-26.
>What is website address, please.
> At 04:55 AM 1/11/01 -0800, you wrote:
> >Dr. Gwen Childs wrote the whole chapter on the subject in 1995: Specimen
> >Preparation for Microscopy and Microanalysis. SM International Press,
> >Chicago, IL.
> >In addition, her www site is loaded with protocols and images.
> >> >Hello Histonetters,
> >> >
> >> >Some of the researchers around here do in situ hybridization followed
> >> >by IHC. However, the harsh conditions of the in situ hybridization
> >> >techniques destroys many of the antigenic sites of interest. Does
> >> >anyone out there use antigen retrieval to recover antigenicity
> >> >following in situ techniques, or does anyone know of any references?
> >> >
> >> >Karen Larison
> >> >University of Oregon
> >>Hi Karen,
> >>ISH techniques sometimes work like "antigen retrieval techniques" for some
> >>antigens. I have used ISH in combination with ICC for pituitary hormones,
> >>and the ICC combined to the ISH was often stronger and with less background
> >>than alone. However, for other antigens, the ISH procedures seem a bit too
> >>harsh, and no antigen retrieval will recover this.
> >>We've tried to combine ISH with ICC for vasopressin, and the immunolabel
> >>was strongly reduced. I suspect that for small peptides like vasopressin,
> >>the proteinase K step combined with the high temperatures of the ISH result
> >>in elutin of some of the peptide.
> >>We have tried microwave antigen retrieval for vasopressin and other
> >>antigens like intermediate filaments, Pit-1, thyroid hormone receptors and
> >>several other antigens, but nothing would do for any of those. I think you
> >>will have to try, but I suspect that antigen retrieval will be of little
> >>help after in situ.
> >> -=-
> >> (o -) O
> >>Paul Klosen, PhD
> >>CNRS UMR 7518 Neurobiologie des Fonctions Rythmiques et Saisonnieres
> >>Universite Louis Pasteur
> >>12 rue de l'Universite
> >>F-67000 Strasbourg, FRANCE
> >>tel: 03.88.35.85.04 fax: 03.88.24.04.61
> >Marek Malecki, M.D., Ph.D.
> >Director and Principal Investigator
> >Molecular Imaging Laboratories, University of California, San Diego
> >address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA92093-0368
> >phone: 8588223373
> >fax: 8584573355
> >pager: 8586161420
> >e.mail: firstname.lastname@example.org
> >e.pager: email@example.com
> >www site: http://mil.ucsd.edu/
> >ftp site: mil1.ucsd.edu
>Veterinary Molecular Biology
>Montana State University
>Bozeman MT 59717-3610
Marek Malecki, M.D., Ph.D.
Director and Principal Investigator
Molecular Imaging Laboratories, University of California, San Diego
address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA92093-0368
www site: http://mil.ucsd.edu/
ftp site: mil1.ucsd.edu
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