Flow cytometric analysis of live human peripheral blood mononuclear cells, untreated (blue) or treated with Cell Stimulation Cocktail (without Protein Transport Inhibitors) (500X) (5 hr; green), using CD69 (FN50) Mouse mAb (PerCP-Cy5.5® Conjugate) #28633 (solid lines) or a concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (PerCP-Cy5.5® Conjugate) #24589 (dashed lines).
1. Prepare the cell suspension in the appropriate cell culture medium.
2. To obtain a 1X final concentration, add the Cell Stimulation Cocktail (without Protein Transport Inhibitors) (500X) directly to the cell suspension at 2 µL/mL.
3. Incubate the cells at 37°C in an atmosphere that contains CO2.
4. Harvest the cells for the immunoassay of interest.
Note: The concentration of cells and incubation time will vary depending on the desired results. This product is an ethanol solution and susceptible to evaporation. Keep tightly sealed when not in use.
Store at -20ºC tightly sealed to prevent evaporation and protected from light. This product is stable for at least 6 months when stored at -20ºC.
Cell Stimulation Cocktail (without Protein Transport Inhibitors) (500X) is an ethanol solution that contains the phorbol ester, PMA (phorbol-12-myristate-13-acetate) and the calcium ionophore, Ionomycin. When added in vitro to cell suspensions, the PMA and Ionomycin activate the cells, leading to increased cytokine production for detection by downstream immunoassays.