Sandwich ELISA analysis of human IL-12 was performed using a Human IL-12 (p70) colorimetric ELISA kit (Product # EHIL12) by loading 50 µL per well of a biotinylated anti-human IL-12 antibody (Product # M121B) in duplicate at 0.03125 µg/mL, and 50 µL per well of Human IL-12 (p70) Recombinant Protein (Product # SIL12) in duplicate at 600, 240, 96, 38.4, 15.4, 0 pg/mL across a 7 µg/mL anti-human IL-12 (Product # M122) pre-coated plate and incubating for 3 hours at room temperature. The plate was washed then incubated with 100 µL per well of Streptavidin-HRP (Product # N504) in all test wells at 1:3,000 for 30 minutes at room temperature. Detection was performed with TMB substrate for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.