BD 555029 GolgiPlug Protein Transport Inhibitor (Containing Brefeldin A) 抑制剂 蛋白转运阻断剂

Stimulation of cells:  Various in vitro methods have been reported for stimulated cells to produce cytokines. Polyclonal activators have been particularly useful for inducing cytokine-producing cells. These activators include the following: concanavalin A, lipopolysaccharide, phorbol esters plus calcium ionophore or ionophore or ionomycin, phytohaemaglutinin, staphylococcus, enterotoxin B, and monoclonal antibodies directed against subunits of the TCR/CD3 complex (with or without antibodies directed against costimulatory receptors, such as CD28).

Procedure for using BD GolgiPlug™:  Add 1 µl of BD GolgiPlug™ for every 1 ml of cell culture (e.g., ~10^6 cells/mL) and mix thoroughly. Treatment of stimulated cells for 4 to 6 hours with BD GolgiPlug™ significantly increases the ability to detect cytokine-producing cells by immunofluorescent staining. It is recommended that BD GolgiPlug™ not be kept in cell culture for longer than 12 hours.

As an alternative to BD GolgiPlug™, investigators may wish to consider using BD GolgiStop™, a protein transport inhibitor containing monensin (Cat. No. 554724). BD GolgiPlug™ and BD GolgiStop™ have been found to have differential effects on intracellular cytokine staining that is time, activator and cytokine dependent. These factors need to be considered when carrying out intracellular cytokine staining.