ANDC DU/Biology Protocols/Seprate Serum

Experiment: To separate serum from blood

Principle:

Blood plasma is the liquid component of blood in which the blood cells are suspended. It makes up about 55% of total blood volume. It is composed of mostly water (90% by volume), and contains dissolved proteins, glucose, clotting factors, mineral ions, hormones, and carbon dioxide (plasma being the main medium for excretory product transportation). Blood plasma is prepared simply by spinning a tube of fresh blood in a centrifuge until the blood cells fall to the bottom of the tube. The blood plasma is then poured or drawn off. Blood serum is blood plasma without fibrinogen or other clotting factors.

Materials required:

1. Glass tubes 2. Glass pasteur pipette 3. Centrifuge (temp 4°C)

Procedure:

1) Collect blood sample (preferably in glass tubes) and leave for 1 hour at 37°C to allow it to clot. 2) Leave sample at 4°C overnight to allow the clot to contract. 3) Using a glass pasteur carefully loosen the clot from the sides of the tube. It is important not to lyse the red cells as they can not then be separated from the serum. 4) Centrifuge the serum at 4000 rpm for 20 minutes at 4°C. 5) Remove the serum from the clot by gently pipetting off into a clean tube using a glass pasteur pipette. 6) Label with the animal number, antigen, total bleed and date. 7) Store the serum at -20°C.

Work in progress, expect frequent changes. Help and feedback is welcome. See discussion page.