How to dilute Brefeldin A?
Formulation: Brefeldin A Solution is supplied as a 1000X in DMSO, which should be diluted to 1X in cell culture medium. Storage: Store the Brefeldin A Solution (1,000X) between 2°C and 8°C. Note: DMSO freezes at this temperature. Recommended Usage: Dilute the 1000X solution to 1X in the tissue culture medium.
Why use Brefeldin A?
Brefeldin A Solution (1000x) is commonly used to enhance intracellular cytokine staining by inhibiting protein transport. This solution causes many of the cytokines to accumulate in the Golgi apparatus and endoplasmic reticulum.
What cellular process does brefeldin a inhibit?
Brefeldin A inhibits vesicle formation and transport between the endoplasmic reticulum and the Golgi apparatus which ultimately results in collapse of the Golgi apparatus into the endoplasmic reticulum via membrane fusion.
What is the difference between GolgiStop and GolgiPlug?
A. The major difference between these two inhibitors is where they work within a cell. BD GolgiStop (monensin) works by accumulation of protein at the endoplasmic reticulum (ER) stage, while BD GolgiPlug (brefeldin) works with accumulation of protein at the golgi complex.
How much does brefeldin cost?
Brefeldin A Solution (1,000X)
| Cat # | Size | Price |
|---|---|---|
| 420601 | 1 mL | $55.00 |
How does PMA ionomycin work?
PMA activates protein kinase C, while ionomycin is a calcium ionophore, and stimulation with these compounds bypasses the T cell membrane receptor complex and will lead to activation of several intracellular signaling pathways, resulting in T cell activation and production of a variety of cytokines.
How does a BFA work?
The blocking of intracellular vesicle movement in BFA-treated cells causes the rapid accumulation of proteins in the ER, thereby disrupting the trafficking of many proteins. BFA inhibits type I interferon (IFN) production by blocking the dissociation of activated STING (stimulator of interferon genes) from the ER.
Is brefeldin a reversible?
Brefeldin A (BFA) is a fungal macrocyclic lactone and a potent, reversible inhibitor of intracellular vesicle formation and protein trafficking between the endoplasmic reticulum (ER) and the Golgi apparatus [1].
What is BFA treatment?
Brefeldin A (BFA) treatment stops secretion and leads to the resorption of much of the Golgi apparatus into the endoplasmic reticulum. This effect is reversible upon washing out the drug, providing a situation for studying Golgi biogenesis.
What is GolgiStop?
The ex vivo addition of BD GolgiStop™, a protein transport inhibitor containing monensin, to in vitro- or in vivo-stimulated lymphoid cells blocks their intracellular protein transport processes. This results in the accumulation of cytokines and/or proteins in the Golgi complex.
How do you dilute monensin?
Monensin is known to be toxic; avoid direct body contact. Ethanol is flammable; keep away from sources of fire. Dilute the 1000X solution to 1X in the tissue culture medium. It is recommended that cells are cultured with monensin for ≤ 24 hours, as this can become toxic for cell viability.
What is PMA stimulation?
What kind of solution is Brefeldin A made of?
Brefeldin A is found naturally as a white to off-white crystalline solid. It forms a clear colorless solution when dissolved. It is soluble in methanol (10 mg/mL), ethanol (5 mg/mL), DMSO (20 mg/mL), acetone, and ethyl acetate (1 mg/mL) without the aid of heating. It is poorly soluble in water (slightly miscible ).
How is the long term effect of brefeldin A determined?
To examine long-term effects of Brefeldin A treatment, cells are seeded at initial concentration 10 5 cells/mL and treated with 0-75 ng/mL Brefeldin A for up to 5 days. At the time indicated, a sample of cells is removed and viable cell number is assessed by standard Trypan Blue exclusion assay.
How much Brefeldin is needed to block growth of hf4.9 cells?
The administration of 25 ng/mL of Brefeldin A completely blocks growth of HF4.9 and HF28RA cells, whereas higher Brefeldin A doses (75 ng/mL) are required to achieve the same effect in HF1A3 cells.
How is Brefeldin A used to treat cancer?
Brefeldin A is a lactone antibiotic and ATPase inhibitor for protein transport with IC50 of 0.2 μM in HCT 116 cells, induces cancer cell differentiation and apoptosis. It could also improve the HDR(homology-directed repair) efficiency and be an enhancer of CRISPR-mediated HDR.