| Enhancement of immunohistochemical staining of scrapie proteins and immune cells within lymph nodes of early scrapie-infected sheep. | |
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MedLine Citation:
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PMID: 21722647 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Transmissible spongiform encephalopathies (TSE) are a group of fatal neurodegenerative diseases that affect animals as well as humans. The oldest of these diseases is Scrapie seen in sheep. Scrapie is caused by an altered form (PrP(sc)), capable of inducing "self-replication" of the normal host prion protein (PrP(c)). There is currently no universal standard for antigen retrieval when using immunohistochemistry to simultaneously stain the PrP(c) protein and other cellular markers. The use of formalin-fixed tissue creates a challenge by concealing the antigenic sites where an antibody would bind, and lengthy antigen retrieval methods must be applied in order to facilitate staining. Further complicating sheep tissue immunohistochemistry is a significant lack of commercial antibodies to sheep cell markers available in research. Here we developed a novel immunohistochemical technique using trypsin, formic acid, and hydrated autoclaving using citraconic anhydride buffer to increase sensitivity of staining for scrapie proteins and immune cell subsets. This allowed us to stain and identify cells within lymphoid tissue associated with early lymphoid pathogenesis in scrapie. |
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Authors:
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Annissa Furr; David Knudsen; Michael B Hildreth; Alan J Young |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2011-06-23 |
Journal Detail:
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Title: Journal of immunological methods Volume: 371 ISSN: 1872-7905 ISO Abbreviation: J. Immunol. Methods Publication Date: 2011 Aug |
Date Detail:
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Created Date: 2011-08-08 Completed Date: 2011-09-27 Revised Date: 2013-05-24 |
Medline Journal Info:
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Nlm Unique ID: 1305440 Medline TA: J Immunol Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 1-7 Citation Subset: IM |
Copyright Information:
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Copyright © 2011 Elsevier B.V. All rights reserved. |
Affiliation:
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Department of Veterinary and Biomedical Sciences, South Dakota State University, Brookings, SD 57007, United States. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antibodies, Monoclonal Deer Formates Humans Immunohistochemistry / methods, veterinary* Lymph Nodes / immunology, metabolism, pathology PrPSc Proteins / immunology*, metabolism* Scrapie / immunology*, metabolism*, pathology Sheep Staining and Labeling / methods Trypsin Wasting Disease, Chronic / immunology, metabolism, pathology |
| Grant Support | |
ID/Acronym/Agency:
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1R15AI072757/AI/NIAID NIH HHS; R15 AI072757-01/AI/NIAID NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Antibodies, Monoclonal; 0/Formates; 0/PrPSc Proteins; 64-18-6/formic acid; EC 3.4.21.4/Trypsin |
| Comments/Corrections | |
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