Isolating plasma from frozen blood samples - (Aug/30/2011 )

Hello

Question. I have frozen blood samples of mice. I need to isolate the blood plasma for protein detection. Does anyone know of a protocol to do this. First I thawed the whole blood sample, which is in EDTA coated vials. I tried spinning down at 3000 rpm for 10' but no clear plasma fraction was present. Also after spinning 14000rpm for 5' no clear fractionation.
Do I need to add something, or did the freezing to -80 already mess up my experiment.

hope you can help

You lysed the blood cells by freezing. You should have spun the samples after collection to obtain the plasma before freezing. You can check your 'detection' method with a couple of samples of known concentrations and run them lysed and non-lysed to determine if your stored samples are of any value.