I want to isolate lymphocytes from rat blood after decapitation and thereafter measure glucocorticoid receptor protein level by western. However, because of experimental design, rats will be killed one after each other with cca 10 min interval in between, cca 10-15 rats per day and there is no posiblity to process samples earlier than after finishing the whole set of rats. The time difference btw first and last blood samples will be cca 5-6 hrs. I wonder how to store the blood for later lymphocyte separation to avoid changes in protein level or whether the GR protein levels could be changed dramatically after keeping the blood samples on ice for 5-6 hrs and then to perform Ficoll gradient centrifugation.
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