16 mL of 2X Lysis Buffer. 2X Lysis Buffer should be diluted 2-fold with deionized or distilled water. Supplementation with protease or phosphatase inhibitors is recommended. You may use RayBiotech Protease Inhibitor Cocktail (AA-PI), RayBiotech Phosphatase Inhibitor Cocktail Set I (AA-PHI-I) or Phosphatase Inhibitor Cocktail Set II (AA-PHI-II), or the RayBiotech Phosphatase / Protease Inhibitor Set (AA-PIPHI). Alternatively, you can purchase inhibitor cocktails from another manufacturer and follow their specifications.