Invitrogen™ eBioscience™ Cell Stimulation Cocktail (plus protein transport inhibitors) (500X)

The Invitrogen™ eBioscience™ Cell Stimulation Cocktail (plus protein transport inhibitors) (500X) is a highly concentrated, ready-to-use reagent formulated to simultaneously stimulate immune cells and retain newly synthesized cytokines intracellularly. The cocktail combines phorbol 12-myristate 13-acetate (PMA) and ionomycin for potent, non-specific cell activation with brefeldin A and monensin, which inhibit intracellular protein transport.

This formulation is widely used for intracellular cytokine staining (ICS) and functional immune profiling by flow cytometry, enabling robust detection of cytokines and other secreted proteins in both in vitro-cultured and ex vivo-isolated cells.

Mechanism of Action

• PMA activates protein kinase C (PKC), triggering downstream signaling pathways that promote immune cell activation.
• Ionomycin increases intracellular calcium levels, synergizing with PMA to induce cytokine production across multiple immune cell types.
• Brefeldin A disrupts ER-to-Golgi transport by inhibiting vesicular trafficking.
• Monensin, an ionophore, further blocks Golgi function by altering ion gradients.

Together, these components induce cytokine production while preventing their secretion, causing intracellular accumulation within the endoplasmic reticulum and Golgi apparatus, which can then be detected by intracellular staining.

Key Features & Advantages

• All-in-one stimulation and inhibition cocktail
• Simultaneous cytokine induction and retention
• Highly concentrated (500X) for economical and flexible use
• Optimized for intracellular cytokine staining (ICS)
• Suitable for both in vitro and ex vivo immune cells
• Reproducible, robust stimulation across many cell types
• Fully compatible with flow cytometry and functional assays

Applications

• Intracellular cytokine staining (ICS)
• Flow cytometric immune profiling
• Functional T-cell and NK-cell assays
• Cytokine production and signaling studies
• Immune activation studies in primary cells
• Cell culture–based functional assays

Reported Applications:

• Flow Cytometric Analysis
• Functional Assays
• Cell Culture

Product Formats & Ordering Information

Technical Specifications

Controls & Related Products

For unstimulated or vehicle control conditions, use the Protein Transport Inhibitor Cocktail (500X) (Cat. No. 00-4980), which contains brefeldin A and monensin without PMA and ionomycin.

Storage & Handling

• Store according to manufacturer’s recommendations
• Thaw and mix gently before use
• Add directly to cell cultures at the recommended dilution
• Use aseptic technique to maintain culture integrity

The Invitrogen™ eBioscience™ Cell Stimulation Cocktail (plus protein transport inhibitors) (500X) is an essential reagent for robust immune cell activation and intracellular cytokine detection. By combining powerful stimulators with protein transport inhibitors in a single formulation, it streamlines workflows and ensures reliable, high-quality results in flow cytometry-based functional immunology studies. It is a trusted solution for researchers investigating cellular immune responses, cytokine signaling, and immune profiling.