Active Noggin (NOG)

NOG (Noggin) is a signaling molecule that plays an important role in promoting somite patterning in the developing embryo, essential for cartilage morphogenesis and joint formation. It is considered as an inhibitor in bone morphogenetic proteins (BMP) signaling, by binding with BMP4, thus a binding ELISA assay was conducted to detect the association of recombinant rat NOG with BMP4. Briefly, NOG were diluted serially in PBS with 0.01%BSA (pH 7.4). Duplicate samples of 100uL were then transferred to BMP4-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-NOG pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37°C. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of NOG with BMP4 was shown in Figure 1 and this effect was in a dose dependent manner.