Materials and Reagents

1. Lympholyte-M (Cedarlane, catalog number: CL5035 )
   
2. RPMI 1640 (Hyclone, catalog number: SH300027.01 ) 
3. Chromium-51 (PerkinElmer, catalog number: NEZ030001MC )
   
4. Poly (I:C) (Sigma-Aldrich, catalog number: P1530 )
   
5. Source of mouse spenocytes: C57Bl/6 mice (Charles Rivers, strain code: 0 27 )
   
6. NK sensitive target cell lines: YAC-1, ATCC, TIB 160
   
7. 1x sterile PBS (Hyclone, catalog number: 21-031-CV )
   
8. Running buffer (Miltenyi Biotech, catalog number: 130-091-221 )
   
9. Washing buffer (Miltenyi Biotech, catalog number: 130-092-987 )
   
10. AutoMACS pro columns (Miltenyi Biotech, catalog number: 130-021-101 )
    
11. AutoMACS buffer (see Recipes)
    
12. Complete RPMI (see Recipes)
    
13. NK cell media (see Recipes)
    

Equipment

1. 50 ml tubes (BD Biosciences, Falcon^®, catalog number: 352098 )
   
2. 15 ml tubes (BD Biosciences, Falcon^®, catalog number: 352096 )
   
3. 96 V well plates (Corning, Costar^®, catalog number: 3894 )
   
4. AutomacsPro Separator (Miltenyi biotech, model: 130-092-545 )
   
5. Gamma counter (PerkinElmer, model: 2470 )
   
6. Incubator (5% CO₂, 37 °C) (Sanyo)
   
7. Centrifuge (when parameters of brakes are unspecified, maxmal acceleration and deceleration are used) (Thermo Fisher Scientific, model: ST40R )
   
8. Dissection instruments (small forceps, scissors)
9. Cell strainers 70 μm (Thermo Fisher Scientific, catalog number: 22363548 )
   
10. DX5 (CD49b) microbeads (Miltenyi Biotech, catalog number: 130-052-501 )
    

Procedure