Figure. The binding activity of GDNF with GFRa2.
Glial cell-derived neurotrophic factor (GDNF) is a small protein that potently promotes the survival of many types of neurons. This protein was shown to promote the survival and differentiation of dopaminergic neurons in culture, and was able to prevent apoptosis of motor neurons induced by axotomy. The most prominent feature of GDNF is its ability to support the survival of dopaminergic and motorneurons. Besides, Glial Cell Line Derived Neurotrophic Factor Receptor Alpha 2 (GFRa2) has been identified as an interactor of GDNF, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse GDNF and recombinant mouse GFRa2. Briefly, GDNF were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to GFRa2-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-GDNF 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℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of GDNF and GFRa2 was shown in Figure 1, and this effect was in a dose dependent manner.