Direct ELISA analysis of Ferritin was performed by coating wells of a 96-well plate with 100ul per well of Human Ferritin or BSA diluted in carbonate/bicarbonate buffer, starting at a concentration of 1 µg/mL and serially diluting 2-fold to a concentration of 2ng/ml, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 100ul per well of a mouse anti-ferritin monoclonal antibody at a concentration of 1 µg/mL for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:25000 for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB substrate for 10 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.