武汉云克隆科技股份有限公司

Neurosecretory protein VGF is specifically expressed in a subpopulation of neuroendocrine cells, and is upregulated by nerve growth factor. The protein shares similarities with the secretogranin/chromogranin family, however, its exact function is not known. Besides, Early Growth Response Protein 1 (EGR1) has been identified as an interactor of VGF, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse VGF and recombinant mouse EGR1. Briefly, EGR1 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to VGF-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti- EGR1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, 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 450/630nm immediately. The binding activity of VGF and EGR1 was shown in Figure 1, the ED50 for this effect is 0.473 ug/mL.