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Brefeldin A (BFA) is a protein transport inhibitor commonly used to enhance intracellular cytokine staining signals by blocking transport processes during cell activation. Especially useful for the intracellular staining of cytokines, BFA leads to the accumulation of most cytokines at the Golgi Complex/Endoplasmic Reticulum (see Jung, et al., 1993). Optimal conditions for use are cell type and time-dependent. Typically, protein transport inhibitors are included during in vitro cell activation cultures for 4-24 hours prior to harvest (see references below for additional information). Brefeldin A Solution is supplied as a 1,000X solution, which should be diluted to 1X in cell culture medium.Product Details
- Brefeldin A Solution is supplied as a 1000X in DMSO, which should be diluted to 1X in cell culture medium.
- 5.0 mg/ml
- Storage & Handling
- Store the Brefeldin A Solution (1,000X) between 2°C and 8°C. Note: DMSO freezes at this temperature.
ICFC - Quality tested
- Recommended Usage
Dilute the 1000X solution to 1X in the tissue culture medium. It is recommended that cells are cultured with brefeldin A for ≤ 24 hours, as this can become toxic for cell viability.
- Application Notes
Completely thaw the solution before use, DMSO freezes at 4°C. We recommend aliquotting the reagent into smaller volumes to avoid repeated freeze-thawing.
(PubMed link indicates BioLegend citation)
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- Product Citations
- Antigen References
1. Current Protocols in Immunology (John Wiley & Sons New York) Unit 6.24 Detection of Intracellular Cytokines by Flow Cytometry (Barbara Foster and Calman Prussin NIAID NIH Bethesda MD).
2. Sander B, et al. 1991. Immunol. Rev. 119:65.
3. Sander B, et al. 1993. J. Immunol. Meth. 166:201.
4. Prussin C, et al. 1995. J. Immunol. Meth. 188:117.
5. Jung T, et al. 1993. J. Immunol. Meth. 159:197.
- Gene ID
- The Brefeldin A solution freezes at 4°C . Is the solution still OK?
We recommend to thaw in warm water after removing from the fridge. The Brefeldin A is dissolved in DMSO, which freezes at 18°C, so this is normal. No one has reported any problems with the freeze/thaw cycles and this is how we recommend it is stored. If customers are concerned, they can aliquot the Brefeldin A to avoid the freeze thaw.
- How do I choose monensin or Brefeldin A solution?
Generally, Brefeldin A is more toxic for longer term incubations, so for shorter stimulations (6 hrs or less) use Brefeldin A. For longer stimulations use monensin. We recommend optimization for each cell type and protocol.