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Article
Enhancement of Immunohistochemical Staining of Scrapie Proteins and Immune Cells within Lymph Nodes of Early Scrapie-infected Sheep
Journal of Immunological Methods
  • Anissa Furr, South Dakota State University
  • David Knudsen, South Dakota State University
  • Michael B. Hildreth, South Dakota State University
  • Alan Young, South Dakota State University
Document Type
Article
Publication Date
8-1-2011
Keywords
  • immunohistochemistry,
  • scrapie,
  • prions,
  • citraconic anhydride,
  • antigen retrieval
Abstract
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 (PrPsc), capable of inducing “self-replication” of the normal host prion protein(PrPc). There is currently no universal standard for antigen retrieval when using immunohistochemistry to simultaneously stain the PrPc 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.
DOI of Published Version
10.1016/j.jim.2011.06.008
Publisher
Elsevier
Rights
Copyright © 2011 Elsevier
Comments

Furr, Annissa, David Knudsen, Michael B. Hildreth, Alan J. Young.2011. Enhancement of immunohistochemical staining of scrapie proteins and immune cells within lymph nodes of early scrapie-infected sheep. J. Immunol. Methods. 371: 1-7. DOI: 10.1016/j.jim.2011.06.008

Citation Information
Anissa Furr, David Knudsen, Michael B. Hildreth and Alan Young. "Enhancement of Immunohistochemical Staining of Scrapie Proteins and Immune Cells within Lymph Nodes of Early Scrapie-infected Sheep" Journal of Immunological Methods Vol. 371 Iss. 1-2 (2011) p. 1 - 7
Available at: http://works.bepress.com/alan_young/95/