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Prion Protein Extraction in Animal Tissues

Instrument: Precellys® 24 Dual

Sample:  Brain or Lymphoid tissues

Protocol:

  • Precellys kits: CK14 (small ceramic beads)
  • Sample : 50 – 150 mg of brain and lymphoid tissues
  • Buffer : glucose 5% (500 - 1500μl) added after grinding
  • Run the Precellys®24 for Brain tissues at 6500rpm, 2x30 sec.,15 sec. break. For Lymphoid tissues at 6500rpm, 3x30 sec., 20 sec. break
  • Prion protein extraction
  • ELISA analysis and Western Blot

Results:

After prion protein extraction, the pathologic protein is detected and quantified by using specific antibodies through an ELISA test, and the pattern of the glycozylated protein is analyzed by Western Blot.

Lanes 4-6 : two positive references
Lanes 1-3 : BSE or Scrapie infected brain tissues
Lanes 5,7,8 : BSE infected lymphoid tissues