Materials and Reagents

1. Fresh or frozen tumor tissues stored at - 80 °C
   
2. Extraction buffer (Life Technologies, catalog number: FNN0011 )
   
3. Antifoam Y-30 Emulsion (Sigma-Aldrich, catalog number: A5758 )
   
4. BCA protein quantification kit (Interchim, catalog number: MP2920 )
   
5. ELISA kits (Peprotech or R&D System)
   
6. Tween-20 (Sigma-Aldrich, catalog number: P-7949 )
   
7. BSA (Sigma-Aldrich, catalog number: A-7030 )
   
8. Avidin-HRP conjugate solution (Sigma-Aldrich, catalog number: A-7419 )
   
9. 10x Dulbecco’s PBS (Life Technologies, Gibco^®, catalog number: 14200-075 )
   
10. ABTS Liquid substrate solution (Sigma-Aldrich, catalog number: A3219 )
    

Equipment

1. Homogenizer such as Precellys (Ozyme BER1011S, France) or ultraturax
   
2. Centrifuge
   
3. 96 wells plates (DUTSCHER SCIENTIFIC, catalog number: 047632 )
   
4. ELISA microplates (Nunc MaxiSorp, catalog number: 439454 )
   
5. Luminoskan (Thermo Fisher Scientific, catalog number: 5210470 )
   

Procedure