04 February 2022 2 2K Report

Hi,

I want to detect Granzyme B after CD8+ stimulation by Dynabeads. My question is :

Ho long should I incubate my Cd8+ with beads to detect (flow cytometry) GzB? and when should I incubate cells with a transport inhibitor cocktail ( Brefeldin A/ Monensine)?

I.e cells should be incubated for 3 days (24 hours could be enough?) with beads and inhibitor cocktail at the same time and then analyzed by GzB intracellular staining? or inhibitor cocktail should be added to the medium 4 hours before Flow cytometry analysis?

Thanks

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