Brefeldin A Monensin
Brefeldin A Monensin - Related Videos
10:15
Identification of the Source of Secreted Proteins in the Kidney by Brefeldin A Injection
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Cited by 3 •
2021
Identifying the cell type responsible for secreting cytokines is necessary to understand the pathobiology of kidney disease. Here, we describe a method to quantitatively stain kidney tissue for cytokines produced by kidney epithelial or interstitial cells using brefeldin A, a secretion inhibitor, and cell-type-specific markers.
05:00
Ex Vivo T Cell Stimulation in the Pancreatic Tumor: A Method to Increase Cytokine Production in T Cells
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2023
This article describes the technique for ex vivo T cell stimulation to increase the intracellular concentration of cytokines such as interferon-gamma or IFN-gamma and tumor necrosis factor-alpha or TNF-alpha in the T cell. These cytokines mediate an effective anti-tumor response and can have a therapeutic role in the...
03:45
Quantification of Inflammatory Mediators in Infected Human Tissue-Derived Cells
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2025
This video demonstrates the quantification of inflammatory mediators during infection by incubating human lung-tissue-derived immune cells with Haemophilus influenzae. Flow cytometry is used to measure cytokine fluorescence in labeled lymphocytes, correlating with the production of pro-inflammatory cytokines in response to bacterial infection.
03:23
An In Vitro Technique to Stimulate Lymphocytes via Pathogenic Bacteria
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2025
This video demonstrates an in vitro technique to stimulate lymphocytes via non-typeable Haemophilus influenzae (NTHi) and to assess the immune response. Incubating isolated peripheral blood mononuclear cells (PBMCs) with NTHi causes the bacterial cells to stimulate T lymphocytes, leading to cytokine production. Upon inhibiting the secretion of cytokines, immunofluorescence staining is performed to detect the accumulated intracellular cytokines in stimulated T lymphocytes.
10:31
Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture
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Cited by 16 •
2016
We describe a protocol to efficiently isolate skin resident T cells from human skin biopsies. This protocol yields sufficient numbers of viable human skin resident lymphocytes for flow cytometric analysis and ex vivo culture.
FAQs
Brefeldin A interferes with activation of ARF1 by targeting guanine nucleotide exchange factors located at the Golgi. Because ARF1 activity supports Golgi membrane organization, its inhibition promotes Golgi disassembly and retrograde movement of Golgi components. This makes the compound useful for examining how cancer cells respond when forward secretory traffic and Golgi architecture are disrupted.
Monensin acts as an ionophore, changing intracellular ion gradients and the pH of the Golgi. These alterations interfere with protein processing and export rather than directly blocking ARF1 activation. Comparing its effects with Brefeldin A helps distinguish consequences linked to Golgi ion and pH balance from those caused by changes in Golgi membrane organization.
The two agents provide complementary perturbations of intracellular trafficking. Brefeldin A primarily tests the importance of ARF1-dependent Golgi organization, whereas monensin challenges ion balance, Golgi pH, processing, and export. If cancer cells respond differently, those differences can indicate whether malignant-cell vulnerability is more closely associated with membrane trafficking, organelle conditions, or both.
Researchers use these pharmacological tools to challenge trafficking and secretory-pathway function in cancer cells, then examine resulting changes in secretion, protein handling, and organelle homeostasis. The responses can expose dependencies that help malignant cells maintain their intracellular organization. Such experiments also support comparisons of how different cancer-cell states tolerate disruption of membrane transport.
A useful analysis focuses on changes in Golgi organization, protein processing and export, secretion, and broader organelle homeostasis. Researchers can also evaluate cellular stress responses and differences in drug sensitivity. Together, these outcomes show how strongly a cancer-cell system relies on intact vesicle trafficking and whether the perturbation produces a selective vulnerability.
Disrupting secretory trafficking can reveal functions that malignant cells depend on for secretion and organelle stability. Differences in sensitivity to Brefeldin A or monensin may therefore identify trafficking-related vulnerabilities and stress responses worth investigating. The findings can guide studies of whether altered vesicle transport or organelle homeostasis might be exploited therapeutically, without assuming that either agent is itself a treatment.