When do you add brefeldin?

Brefeldin A and/or monensin is added at the time of stimulation (for peptides) or after 2 h (for proteins, to allow for intracellular antigen processing, which is compromised by the secretion inhibitor).

How do you dilute brefeldin A?

Brefeldin A Solution is supplied as a 1000X in DMSO, which should be diluted to 1X in cell culture medium. Store the Brefeldin A Solution (1,000X) between 2°C and 8°C. Note: DMSO freezes at this temperature. Dilute the 1000X solution to 1X in the tissue culture medium.

What is brefeldin a used for?

Brefeldin A (BFA) is a protein transport inhibitor commonly used to enhance intracellular cytokine staining signals by blocking transport processes during cell activation.

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.

What is ELISpot used for?

Enzyme-linked immunosorbent spot (ELISpot) is a highly flexible assay and can be adapted to multiple readout formats. ELISpot assays are quantitative and measure key cellular functions of immune system cells. ELISpot has been used to assess both adaptive and innate immune responses.

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].

Which drugs causes the dissolution of the Golgi apparatus with ER?

Mechanism of action 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 Golgi stop?

Description. 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.

What is ionomycin used for?

Ionomycin is a membrane permeable calcium ionophore produced by the bacterium Streptomyces conglobatus. This ionophore facilitates the transfer of calcium ions (Ca2+) into and out of cells [1]. Ionomycin can be used to increase intracellular calcium levels triggering cell death through apoptosis and autophagy [1].

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