Materials and Reagents

1. Mice
   
2. DNase I (100 mg/ml) (Sigma-Aldrich, catalog number: DN25 )
   
3. 1x PBS
   
4. HEPES (Life Technologies, Gibco^®, catalog number: 15630-080 )
   
5. Magnesium chloride solution (Sigma-Aldrich, catalog number: M1028 )
   
6. Potassium chloride solution (Sigma-Aldrich, catalog number: 60121 )
   
7. Nonidet P40 (NP-40) (Roche Diagnostics, catalog number: 11332473001 )
   
8. Fetal calf serum (FCS)
   
9. Bovine serum albumin (BSA)
   
10. 0.25% trypsin (Life Technologies, Gibco^®, catalog number: 15050 )
    
11. CD45 MicroBeads kit (Miltenyi Biotec, catalog number: 130-052-301 )
    
12. Cytoplasmic extract buffer (see Recipes)
    
13. Nuclear extract buffer (see Recipes)
    

Equipment

1. Tweezers and scissors
   
2. 26 G1/2 needles (BD, catalog number: 309659 )
   
3. Cell strainer (100 μm and 40 μm) (BD Biosciences, Falcon^®, catalog numbers: 352360 and 352340 )
   
4. 10 ml and 1 ml syringe (BD, catalog numbers: 309604 and 305111 )
   
5. Filter tips (Eppendorf)
   
6. 15 ml Corning tubes (Corning)
   
7. DNase, RNase-free Eppendorf tubes (Eppendorf)
   
8. Tissue culture dishes (60 x 15 mm style) (BD Biosciences, Falcon^®, catalog number: 353002 )
   
9. MACS device (Miltenyi Biotec, model: 0 16210 )
   

Procedure