Prion Protein MAb Ascites IHFG IgG1 Isotype

Recognizes a conserved epitope (IHFG) on the prion protein in tissues from sheep, cattle, mule deer, elk, white tailed deer and humans. IgG1 isotype. Suitable for detecting agents of TSEs in ruminant species. Techniques include immunoassasys of formalin-fixes tissues, Western immunoblot, immunohistochemistry (except in cervid lymphoid tissue – F99/97.6.1), and ELISA. F89-160.1.5 (Lot F89-008) was evaluated by immunohistochemistry (IHC) of brain and lymph node from a scrapie-infected sheep, and brain and lymph node from a sheep with no known exposure to scrapie. The antibody was diluted to 0.5 µg/ml and incubated for 30 minutes with pretreated tissues. Dectection was performed on a Ventana immunostainer using biotinylated goat anti-mouse Ig. Streptavidin-horseradish peroxidase and AEC as the chromogen/substrate. There was no staining of the tissue form the sheep with no exposure to scrapie. Staining of the brain and lymph node of the scrapie-infected sheep was intense. The quality of this lot of antibody is excellent. Tissues for IHC with this antibody are paraffin-embedded formalin-fixed tissues, often decontaminated by incubation of formalin-fixed tissue in formic acid for 60 minutes before re-equilibration in formalin and routine processing and embedding. Three to 5 micron sections are rehydrated, Pretreated first with acid (98% formic acid 5-20 minutes, then rinsed and neutralized with several changes of 0.1 M Tris buffer, pH 7.5) then with heat (autoclaved 121°C for 20 minutes in 0.1 M Tris buffer, Ph 7.5 or in modified citrate buffer, pH 6.1, such as DAKO Target Retrieval Buffer).