Hi everyone,
I would like to collect some serum from my whole blood sample to test performing an ELISA. How much calcium should be added to clot my blood that has been collected in a tube containing EDTA
Go to 10 -15 mM and you are on the safe side in case of "underfilling". of the tube which is not uncommon.
Or add a trace of batroxobin. It generated fibrin quite specifically.
Or perform the ELISA directly from plasma. In many cases this will work as well. Thi would be the simpler way to solve the problem.
Your best option is to work with plasma as long as you are not measuring electrolytes. However, you may have to overpower the EDTA with much calcium as suggested but this may affect the morphologies of the cells and other parameters you may wish to measure.
You cannot test clotting in a serum sample, as serum is plasma without the clotting factors. If you wish to test clotting you need a sample still containing these.
Further EDTA is not a very good anticoagulant if you wish to test clotting as this might disrupt some of the clotting factors. You might be able to clot the sample, but I would not be completely convinced that the results are reliable.
I suggest you collect your blood in citrate instead of EDTA and run the test with plasma.
Anna, I'm not testing clotting, I want to clot the blood to obtain serum
Bill
Can someone suggest labs where my graduate students can have hand-on experience on Microparticle studies?
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