Cell Stimulation Cocktail (with Protein Transport Inhibitors) (500×) - sterile-filtered,BioReagent,500×

Cat. No.: C1508366

GRADE & PURITY Sterile-filtered, BioReagent, 500×

Synonyms

Cell Stimulation Cocktail (Protein Transport Inhibitors Plus, 500×) | Cell Stimulation Cocktail (with Protein Transport Inhibitors) (500X) | Cell Activation Cocktail (with Brefeldin A) | Cell Stimulation Cocktail (plus protein transport inhibitors) (500X)

Storage

Protected from light,Store at -20°C

Shipped In

Ice chest + Ice pads

Application

Flow Cytometry

Descripción general

This product is a complete “stimulate-and-block” cocktail provided as a  500× concentrate. In addition to the classic stimulators PMA (Phorbol ester) and Ionomycin, it contains highly effective protein transport inhibitors Brefeldin A and Monensin Sodium. This formulation not only robustly activates immune cells (e.g., T cells, NK cells) to induce cytokine synthesis but also efficiently blocks Golgi-mediated transport, leading to the potent intracellular accumulation of cytokines (e.g., IFN-γ, TNF-α, IL-2). It is a ready-to-use solution optimized specifically for Intracellular Cytokine Staining (ICS) analyzed by flow cytometry.

Key Components:

• Ethonal
• PMA 25μg/mL， lonomycin 500μg/mL，brefeldin A 1.5mg/mL，Monensin sodium salt 0.7mg/mL

Applications

• Standardized stimulation and blockade for Intracellular Cytokine Staining (ICS) and flow cytometry.
• Detection of cytokine-producing capacity in immune cells (e.g., antigen-specific T cells) at the single-cell level.
• Simultaneous analysis of cell surface markers and intracellular functional molecules.
• NOT suitable for experiments requiring supernatant collection for ELISA, ELISpot, or multiplex assays (please use the version without protein transport inhibitors, Cat# C1508365 ).

Instructions for Use

Standard Stimulation Protocol (For ICS)

• Cell Preparation: Suspend target cells (e.g., human PBMCs, mouse splenocytes) in complete medium at an appropriate density (recommended 1-2×10^6 cells/mL) and aliquot into culture plates or FACS tubes.
• Stimulation: Add this cocktail to the cell suspension at a 1:500 ratio. For example, add 2 µL of the 500X concentrate to 1 mL of cell suspension. Mix gently.
• Incubation (Stimulation & Blockade): Incubate cells in a 37°C, 5% CO₂ incubator. Critical Incubation Time: 4 to 6 hours. This duration ensures sufficient cytokine induction and optimal intracellular accumulation with minimal impact on cell viability, representing the ideal window for ICS.
• Cell Processing: After stimulation, proceed directly to surface staining, followed by fixation, permeabilization, and intracellular staining.

Key Precautions

• This cocktail contains optimal concentrations of both stimulators and inhibitors. Do NOT add any other protein transport inhibitors.

• Stimulation time is critical. 4-6 hours is recommended. Shorter times may yield insufficient cytokine, while longer times (>6 hours) may increase cytotoxicity and background.

• Because secretion is completely blocked, the supernatant from cells stimulated with this product CANNOT be used for soluble cytokine detection (e.g., ELISA).

• Aliquot upon receipt to minimize freeze-thaw cycles.

Specifications

Sinónimos

Cell Stimulation Cocktail (Protein Transport Inhibitors Plus, 500×) | Cell Stimulation Cocktail (with Protein Transport Inhibitors) (500X) | Cell Activation Cocktail (with Brefeldin A) | Cell Stimulation Cocktail (plus protein transport inhibitors) (500X)

Especificaciones y pureza

sterile-filtered, BioReagent, 500×

Estabilidad y almacenamiento

Aliquot upon receipt and store frozen at -20°C(12 months). Protect from light. Avoid repeated freeze-thaw cycles

Condiciones de almacenamiento de almacenamiento

Protected from light, Store at -20°C

Enviado en

Ice chest + Ice pads

Este producto requiere envío en cadena de frío. Los servicios terrestres y otros servicios económicos no están disponibles.

Grado

BioReagent, Sterile-filtered