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intracellular cytokines

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Posted 12 September 2009 - 06:09 AM


I am searching for a method for intracellular cytokine staining. So far I have a few options:

-stimulate cells for 6 hours, stain (surface markers and intracellular)
-stimulate cells for 6 hours, store in fridge o/n, stain next day
-stimulate cells overnight, stain next day

Can anyone advise which is the best way to go? do you typically stimulate and stain in a 96 well plate or in falcon tubes?
If i am to incubate the cells o/n in golgi plug and stimulant (pma/iono), should I reduce the concentrations?
What are the optimal concentrations of golgi plug amd pma/iono?

Thanks in advance,


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