Indirect ELISA was performed by coating BSA-Estradiol ~30 ng/well (by Estradiol amount), followed by blocking with 1% BSA. Serial diluted Estradiol antibody 68600-1-Ig was added to the plates and incubated at 37℃. HRP-Goat anti-mouse was used for detection (top X-right Y, blue curve). Competitive ELISA was performed similarly except that different concentration of Estradiol was mixed in 30ng/mL primary antibody (bottom X-left Y, red curve). This data was developed using the same antibody clone with 68600-1-PBS in a different storage buffer formulation.

BSA-Estradiol was coated in 96 well plate at ~30 ng/well (by Estradiol amount), followed by blocking with 1% BSA. Different concentrations of Estradiol standard as well as diluted urine from different donor were mixed with 30 ng/mL Estradiol antibody 68600-1-Ig respectively. Urine Estradiol was calculated based on standard curve. Please notice that the level from different donor might be different between individuals. This data was developed using the same antibody clone with 68600-1-PBS in a different storage buffer formulation.