IUBio

[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.

Regards.

Marcella Sarzotti-Kelsoe


METHOD

The method for clotting plasma will depend on the anticoagulant. The
following will work for donor plasma anticoagulated with ACD, CPD or
CPD-A1.

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

Method:
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.

Reference:
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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