Anti-Denosumab Antibody (16F2), mAb, Mouse (GenScript, A02230) blocks Denosumab binding with Human RANKL recombinant protein.
Coating antigen: Denosumab 1 µg/ml.
RANKL final concentration: 1.2 ng/ml.
Anti-Denosumab antibody dilutions start from 10 μg/ml.
IC₅₀= 0.11 µg/ml.

ELISA binding of Anti-Denosumab Antibody (16F2), mAb, Mouse (GenScript, A02230) with Denosumab. While the antibody does not recognize the human IgG (data not shown).
Coating antigen: Denosumab, 1 µg/ml.
Anti-Denosumab Antibody (16F2), mAb, Mouse (GenScript, A02230) dilution start from 1,000 ng/ml.
EC₅₀= 20.03 ng/ml.

Standard curve of Denosumab Sandwich ELISA by anti-idiotype antibodies in cynomolgus monkey serum.

In this ELISA assay, Anti-Denosumab Antibody (3B1), mAb, Mouse (GenScript, A02229) was coated at a concentration of 1 μg/ml, and Anti-Denosumab Antibody (16F2), mAb, Mouse (GenScript, A02230) conjugated with Biotin was used as a detection antibody at a concentration of 0.5 μg/ml.

In this ELISA assay, a four-parameter logistic curve fitting program was used to create a standard curve with the R-Square equal to 0.99987. The denosumab sdandard was diluted with fresh cynomolgus monkey serum with a volume ratio of 1:10. The sensitivity of detecting Denosumab is up to 1.56 ng/ml.

In this ELISA assay, the OD450 values at each concentration measured in different diluted serum matrix were not significantly different from those of the dilution buffer. The signals from 10 cynomolgus monkey serum samples diluted 10 times or more approached those of the dilution buffer to set an MRD of 10 to reach a concentration within the dynamic range, without altering the assay precision or accuracy.