[Immunology] Plasma to serum protocols?

Marcella Sarzotti Kelsoe, Ph.D. via immuno%40net.bio.net (by marcella.sarzottikelsoe from duke.edu)
Wed May 29 17:14:01 EST 2013

To Whom it may concern,

I found the following method on Google and it directed me to your email. I would like to know more about this protocol but I do not have access to the book.

 - Is there a published paper that describes this method in more details?

 - Are both CaCl2 and Thrombin necessary?

 - Can I use human instead of bovine thrombin?

 - Can I keep the sample in the fridge overnight, instead of freezing it.

 - Does this process affect/alter antibodies and their function?

I would really appreciate it if you could answer my questions or direct me to someone that could.


Marcella Sarzotti-Kelsoe


The method for clotting plasma will depend on the anticoagulant. The
following will work for donor plasma anticoagulated with ACD, CPD or

Bovine thrombin 500 units/ml
Calcium chloride 10%wt/vol

Add 0.25 ml CaCl2 per 100 ml plasma.
Add 0.2-0.4 ml thrombin per ml plasma
Mix well and allow to clot at room temp. for at least 4 hours.
Freeze converted plasma at -20C overnight.
Thaw at room temp.
Centrifuge at 5000g for at least 5 min.
Remove serum and store at -20C until required.

W. John Judd. Methods in Immunohematology. 2nd. Ed., Mongomery Scientific
Publications. Durham, NC.

Robertson D. Davenport, MD
Assistant Professor                    Associate Medical Director
Department of Pathology                Blood Bank and Transfusion Service
University of Michigan Medical School  University of Michigan Hospitals
rddvnprt at umich.edu<http://iubio.bio.indiana.edu/biomail/listinfo/immuno>

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