Google Groups no longer supports new Usenet posts or subscriptions. Historical content remains viewable.
Dismiss
Plasma to serum protocols?
2 views
Skip to first unread message
Marcella Sarzotti Kelsoe, Ph.D.
May 29, 2013, 6:14:01 PM5/29/13
to imm...@magpie.bio.indiana.edu, Lily Daniell, Ph.D.
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>
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>
0 new messages