Intracellular Cytokine (INF-gamma) Staining Assay

Materials and Reagents

1. PE Rat Anti-Mouse IFN-γ (BD Biosciences, catalog number: 554412 )
   
2. PE Rat IgG1 κ Isotype Control (BD Biosciences, catalog number: 554685 )
   Note: The above antibodies have been tested by the author and may be substituted with the antibodies desired by users.
   
3. BD Cytofix/Cytoperm^TM Fixation/Permeabilization Solution Kit with BD GolgiStop^TM (BD Biosciences, catalog number: 554715 )
   
4. 1x phosphate buffered saline (PBS)
   
5. FACS staining buffer (1x PBS, 2% FBS)
   
6. Phorbol 12-myristate 13-acetate (PMA) (Sigma-Aldrich, catalog number: 79346 )
   
7. Ionomycin (Sigma-Aldrich, catalog number: I9657 )
   

Equipment

1. BECKMAN centrifuges and rotor (Beckman Coulter)
   
2. Incubator
   
3. 96 well plates
   
4. Falcon round-bottom tubes
   

Procedure