Active Sex Hormone Binding Globulin (SHBG)

Sex hormone-binding globulin (SHBG) or sex steroid-binding globulin (SSBG) is a glycoprotein, a molecule that consists of a carbohydrate plus a protein, and is produced mainly in your liver. It binds to three sex hormones found in both men and women: estrogen, dihydrotestosterone (DHT) and testosterone. Its job is to transport these hormones throughout your blood to other tissues in your body. Thus a binding ELISA assay was conducted to detect the interaction of recombinant human SHBG and bsa-conjugated testosterone(Testo) . Briefly, biotin-linked recombinant human SHBG were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to bsa-conjugated Testo -coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with HRP conjugage for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of SHBG and Testo was shown in Figure 1, and this effect was in a dose dependent manner.