I took blood from mouse tails for serum collection and left them in a temperature controlled centrifuge at 4c overnight. The serum I collected was pink-tinged, but still fairly clear. I would like to use this serum to measure serum insulin levels, FFAs, and TGs, but I'm not sure how this error in processing might affect this data. Although the layers were separated, I'm assuming there will degradation products from RBCs and leukocytes, and potentially release of intracellular proteases. However, since it was temperature controlled I'm optimistic that the analysis might still be possible and analyzable.
What factors might I want to consider when considering the integrity of this sample for my desired analyses?
Its always a good and reliable practice to separate the serum from RBCs for storage , because RBCs will continue hemolyzing as the time goes on and the serum might turn pink-red which may interfere with colorimetric assays.
low temperature will delay the mentioned processes..
Dear Samir
some extent of hemolysis may be accepted during the analysis of ELISA based /enzymatic analysis. I hope that your results would not be changed. I suggest you, analyze the tests by following same SOP in both control and test groups. It will equalize your interference (if any) in both the group that will not change the outcome of experiment.
It's a good thing you left it a temperature-controlled centrifuge. It will slow down the rate of hemolysis
If the proper anticoagulant lithium heparin is in the bottle the serum is collected. I believe it can still be used after 12hrs
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