GRADE & PURITYSterile-filtered?Sterile-filtered — passed through a 0.2µm filter to remove microorganisms. Use as a ready aseptic solution for culture and sensitive applications.BioReagent?BioReagent grade — tested suitable for life-science and molecular-biology use. Use for cell culture, assays, and biochemical work needing biological compatibility.500×
sterile-filtered,BioReagent,500× BioReagent,Sterile-filtered for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
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Storage & shipping
Protected from light,Store at -20°C Ships Ice chest + Ice pads Check lot-specific COA for exact specifications.
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Quality documents
SDS, COA, datasheet, and spec sheet available for download. Lot-specific COA accessible via lot number lookup.
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Literature proof
Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
Overview
Product Introduction
This product is a classic 500X concentrated cell stimulation cocktail containing an optimized ratio of Phorbol 12-myristate 13-acetate (PMA) and Ionomycin. The synergistic action of these components provides robust activation of immune cells such as T cells and NK cells, inducing gene expression and natural secretion of multiple cytokines (e.g., IFN-γ, TNF-α, IL-2). This cocktail does NOT contain protein transport inhibitors like Brefeldin A or Monensin, making it ideal for studying natural cytokine secretion kinetics, signal transduction pathways, and assays requiring supernatant collection.
Key Components:
Ethonal
PMA 25μg/mL, lonomycin 500μg/mL
Applications
Immune cell activation for ELISA, ELISpot, or multiplex immunoassays (requires supernatant analysis).
Functional assessment of immune cells (e.g., T cell activation, proliferation).
Cell signaling studies (PKC and calcium signaling pathways).
Important: For intracellular cytokine staining and flow cytometry analysis, protein transport inhibitors MUST be added separately during stimulation (please use the version with protein transport inhibitors, Cat# C1508366).
Instructions for Use
Standard Stimulation Protocol
Dilution: Dilute the cocktail 1:500 in complete culture medium immediately before use. For example, add 2 µL of the cocktail to 1 mL of cell suspension.
Stimulation: Incubate cells in a 37°C, 5% CO₂ incubator.
For secretion/gene expression assays: A 4-6 hour stimulation is recommended.
For long-term stimulation/supernatant collection: Can be extended to 16-48 hours (please optimize the duration).
Key Precautions
Optimization of stimulation time (4-48 hours) is recommended for first-time use or new cell systems.
Prolonged stimulation (>24 hours) may affect cell viability and phenotype. Pre-experiment evaluation is advised.
This product lacks protein transport inhibitors. For ICS, inhibitors must be added for the final 4-6 hours of stimulation.
Aliquot upon receipt to minimize freeze-thaw cycles.
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Products are supplied to verified businesses, institutions, and qualified professionals for research and development use only. Not for use in humans, animals, diagnosis, or therapy.