Mouse Glypican 4 protein, His tag

Glypican 4 (GPC4) is a cell surface heparan sulfate proteoglycans which composed of a membrane-associated protein core substituted with a variable number of heparan sulfate chains. Members of the glypican-related integral membrane proteoglycan family (GRIPS) contain a core protein anchored to the cytoplasmic membrane via a glycosyl phosphatidylinositol linkage. These proteins may play a role in the control of cell division and growth regulation. Besides, Fibroblast Growth Factor 2, Basic (FGF2) has been identified as an interactor of GPC4, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse GPC4 and recombinant mouse FGF2. Briefly, GPC4 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to FGF2-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-GPC4 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 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of of GPC4 and FGF2 was in a dose dependent manner.