A mass of 175 mg ± 20 mg of frozen brain tissue was homogenized to a concentration of 10% (wt/vol) in 5% glucose in distilled water in grinding tubes (Bio-Rad), using a TeSeE Precess 48 homogenizer (Bio-Rad). The presence of PrPres in transgenic mice brains was determined by means of WB analysis, as described elsewhere [5 (link), 27 (link)]. Ten to 100 μL of a 10% (wt/vol) brain homogenate was digested with Proteinase K, loaded on 12% Bis-Tris Gel (Criterion XT; Bio-Rad), and detected with Sha31 monoclonal antibody [28 (link)].
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Detection of PrP Res in Transgenic Mouse Brains
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Corresponding Organization :
Other organizations : Centre de Recerca en Sanitat Animal, University of California, San Diego, Norwegian Veterinary Institute, École Nationale Vétérinaire de Toulouse, Institut National Polytechnique de Toulouse
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Variable analysis
independent variables
- Mass of frozen brain tissue
- Presence of PrP^res in transgenic mice brains
- Concentration of brain homogenate (10% wt/vol)
- Diluent (5% glucose in distilled water)
- Homogenization method (TeSeE Precess 48 homogenizer)
- Proteinase K digestion
- Gel electrophoresis (12% Bis-Tris Gel)
- Detection method (Sha31 monoclonal antibody)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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