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hCG / Human Chorionic Gonadotropin Antibody (MIH9827) in ELISA
Direct ELISA analysis of hCG proteins was performed by coating wells of a plate with a purified hCG protein extracted from human urine at a concentration of 10ug/ml overnight at 4C. The plate was washed 3 times with ELISA Wash Buffer (Product ## N503) and blocked with 300 ul of StartingBlock (PBS) Blocking Buffer (Product ## 37538) for at least 1 hour at room temperature. After washing 100ul of hCG monoclonal antibody (Product ## MIH9827) was added to wells in duplicate at 10, 5, 2.5, 1.25, 0.62, 0.31, 0.15, 9.5, 0.078, 0.039, 0.019 and 0 ug/ml concentrations, and the samples were incubated for 2 hours at room temperature. The plate was washed, and then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG secondary antibody (Product ## 31430) at a dilution of 1:10,000 for 1 hour at room temperature, and washed again with ELISA Wash Buffer. The plate was developed by incubating 100ul per well of 1-Step Ultra TMB substrate (Product ## 34028) per well for 10 minutes at room temperature in the dark. The reaction was stopped with 100ul per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550nm.
