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. The increased accumulation of cytokines in the cell enhances the detectability of cytokine-producing cells with flow cytometric analysis.
Investigators should note that the appearance of BD GolgiStop™ may range in color from clear (colorless) to a light yellow.
Suggested Companion Products
BD Cytofix/Cytoperm Plus Kit (with BD GolgiPlug) RUO 250TestsCat No: 555028
BD Cytofix/Cytoperm Plus Kit (with BD GolgiStop) RUO 250TestsCat No: 554715
Protein Transport Inhibitor (Containing Brefeldin A) RUO 1mLCat No: 555029
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Preparation and Storage
Store undiluted at 4°C.
Product Notices
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Stimulation of Cells:Various in vitro methods have been reported for stimulating 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 ionomycin, phytohaemaglutinin, staphlylococcus, entertoxin 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 GolgiStop™: Add 4 µl of BD GolgiStop™ for every 6 mL of cell culture (e.g., ~10^6 cells/mL) and mix thoroughly. Treatment of stimulated cells for 4 to 6 hours with BD GolgiStop™ significantly increases the ability to detect cytokine-producing cells by immunofluorescent staining. It is recommended that BD GolgiStop™ not be kept in cell culture for longer than 12 hours.
As an alternative to BD GolgiStop™, investigators may wish to use BD GolgiPlug™ , a protein transport inhibitor containing brefeldin A (Cat. No. 555029).BD GolgiStop™ and BD GolgiPlug™ have been found to have differential effects on intracellular cytokine staining that is time, activator and cytokine dependent. These factors must be considered when carrying out intracellular staining.
Danger:BD GolgiStop™ Protein Transport Inhibitor, containing monensin (component 51-2092KZ) contains 99.61% ethanol (w/w) and 0.26% monensin, mononatriumsalz (w/w).
Hazard statements:
Highly flammable liquid and vapor.
Causes serious eye irritation.
Harmful if swallowed.
Precautionary statements:
Keep away from heat/sparks/open flames/hot surfaces.No smoking.
Wear protective gloves / eye protection.
Wear protective clothing.
IF ON SKIN (or hair): Remove / Take off immediately all contaminated clothing.Rinse skin with water / shower.
IF IN EYES: Rinse cautiously with water for several minutes.Remove contact lenses, if present and easy to do. Continue rinsing.
IF SWALLOWED: Call a POISON CENTRE/doctor if you feel unwell. Rinse mouth.
Dispose of contents / container in accordance with local / regional / national / international regulations.Keep container tightly closed.
