Semaglutide Antibody

Indirect ELISA analysis of Semaglutide was performed by coating wells of a 96-well plate with 100 µl per well of GLP-1(7-36) antigen (HY-P0054A, Red), GLP-1(7-36) antigen (HY-P1145, Blue), Semaglutide antigen (HY-114118, Black), Tirzepatide antigen (HY-P1731, Green) at a concentration of 1 µg/mL overnight at 4 °C. Wells of the plate were washed and blocked with 1% BSA in 1× PBST buffer. The plate was then incubated with 100 µl per well of Semaglutide antibody (HY-P810528), starting at an initial concentration of 10,000 ng/mL and serially diluted down to 0.01 ng/mL, for 2 hours at room temperature. The plate was washed and incubated with 100 µl per well of an HRP-conjugated goat anti-rabbit IgG secondary antibody (HY-P8001) at a dilution of 1:10,000 for one hour at room temperature. Detection was performed using TMB substrate for 8 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid, and absorbances were read on a spectrophotometer at 450 nm.