BD Biosciences
Protein Transport Inhibitor (Containing Monensin)
554724
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As an immunology lab, we routinely assess intracellular cytokine production from T cells and other immune cells. We stimulated cells using PMA/Ionomycin to maximize cytokine production, but we also use a protein transport inhibitor to block secretion in order to analyze cytokines using flow cytometry. We use GolgiStop as a more potent inhibitor compared to other inhibitors for a short stim.
Flow Cytometry Analysis of Cytokines
Splenocytes or T cell cultures
Prepare stimulation media containing PMA/Ionomycin along with GolgiStop. Resuspend T cells in this solution for 4 hours before processing for flow cytometry of intracellular cytokines.
Determine the correct amount of time for the appropriate cytokines.
Using flow cytometry, we are easily able to assess IFNg secretion from T cells by blocking transport in CD8 T cells during stimulation.
N/A
Easy to use. Is not frozen so no need to thaw
Can be toxic to cells if cultured for prolonged periods of time
None really. Easy to use and widely used by the immunology scientific community.