VMC 221: Veterinary Immunology and Serology (1+1)

PROCEDURE

• A saturated solution of ammonium sulphate (760g/liter) is made by dissolving the salt in hot distilled water and cooling to the room temperature.
• pH of the clear supernatant is adjusted to 7.0 using ammonia solution.
• For precipitation 40 ml of saturated ammonium sulphate solution is added gradually to 60 ml of serum under constant stirring at room temperature.
• The resultant suspension is allowed to stand for 30 minutes with occasional stirring.
• The precipitate is collected by centrifugation at 3000 rpm for 20-30 minutes
• The precipitate is dissolved in 30 ml of distilled water and precipitated again by gradual addition of 20ml of saturated ammonium sulphate solution and the cycle is repeated twice.
• The final precipitate is dissolved in 15-20 ml of 0.15 M sodium chloride and ammonium sulphate is removed by dialysis.

For other procedures refer...