Skip to content
Medicines & Healthcare products Regulatory Agency
The National Institute for Biological Standards and Control

Confidence in biological medicines

  • Stay connected
  • Shopping Basket
  • Pay Now
  • Login / Register
  • Home
  • Products
  • Standardisation
  • Control testing
  • Science and research
  • Expert services
  • About us
  • Advanced therapies
  • Virology
  • Bacteriology
  • Biotherapeutics
  • Analytical sciences
  • Diagnostics
  • A to Z listing
  • Publications
  • Home  /  
  • Science and research  /  
  • Virology  /  
  • CJD resource centre  /  
  • Available samples  /  
  • WHO Reference Reagents  /  
  • NHBX0 - 0001  /  
  • Brain Tissue Preparation
 

Preparation of Infectious CJD Standard for Transmission and Biochemical Studies

Equipment :


Eppendorf EDOS 5222 Liquid dipensing system


CamLab Omni-Mixer ES Homogeniser


Class 2 Microbiological Safety Cabinet

 

Solution


0.25M-sucrose (MW 342.30) : 85.56g/L in pyrogen-free water.
Pre-chilled to ~4 degrees centigrade

 

Procedure:

 

  1. Weigh frozen brain. Thaw enough to allow slicing into 4-5g chunks and transfer to homogenisation container.
  2. Homogenise approx. 20g and 180 mL sucrose solution in sealed 250 mLcontainer using programmable Omni-Mixer. Setting at 4000rpm, 6 x 30seconds.*
  3. Transfer homogenate to 2L beaker on ice/salt. Stir on magnetic stirrer.
  4. Repeat homogenisation : 4 x 20g brain to create a pool of 1L of 10%-brain homogenate.
  5. Dispense 100 x 0.5 mL into pre-labelled, Nalgene 1.2mL cryovials, immerse in bleach (10%-Chloros) for >15 minutes at 4oC.
  6. Rinse in water to remove surface bleach. "Flash Freeze " by immersing in liquid nitrogen for 5 minutes.
  7. Transfer onto dry ice for transport to storage facility at NIBSC (-80 oC).

 

* Shown to be adequate by facsimile homogenisation of frozen sheep brain

  • Careers
  • Terms and conditions
  • Accessibility
  • Privacy notice
  • Cookies
  • Sitemap