Thermo Fisher Scientific
Brefeldin A Solution (1000X)
00-4506-51
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Cells were incubated for 5 hours in a stimulation medium consisting on PMA and Ionomycin in order to stimulate cytokine production. Brefeldin A solution was added at T=0 hr in order to stop the cytokines from being exported out of the cell, therefore building up the intracellular concentration of cytokines to levels that can be detected via flow cytometry.
Flow Cytometry
Mouse spleenic and lymph node cells
400K T cells were incubated with 20ng/ml PMA, 1microM ionomycin, and 4microL of 1000X Brefeldin A in 1mL of culture medium for 5 hours. Post incubation cells were collected and examined via flow cytometric analysis for the presence of intracellular cytokine production.
Adding Brefeldin A in higher concentrations does not increase intracellular yield after 5 hours.
The 100X Brefeldin A works very well to stop the trafficking of intracellular cytokines outside of the cell.
N/A
It is inexpensive and works at low concentrations
If you leave Brefeldin A in culture for too long it will cause cellular death
This is a great reagent to use if you want to study intracellular cytokine production.