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Cell Stimulation for Cytokine Production Protocol

Posted by Michaela Frister on Aug 21, 2014 10:09:02 AM

Cell Stimulation Cocktail (500X) Cat. No. TNB-­4975-­UL100

Note: Cell Stimulation Cocktail is supplied as a 500X solution in Ethanol. Product should be stored at ­20°C, protected from light and tightly sealed to prevent evaporation.

Other Materials Required

  • RPMI complete medium (RPMI with 10% FBS, 1% pen/strep, 50 µM 2­ME)

For activation of mouse splenocytes:

  1. Harvest spleen and prepare single cell suspension in RPMI complete medium at approximately 5­-10 x 10^6 cells/mL.
  2. Add 1X (2 µL/mL) Cell Stimulation Cocktail (500X) directly to the cell suspension.
  3. Culture overnight (6-18 hours) at 37°C, 5% CO2 . We typically leave this culture overnight which will generate a nice population of IL­2 and IFN­gamma expressing cells.

Download PDF here!

Topics: cytokine, protocol, stimulation, support

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