Mouse igf1 elisa kit

VEGF-A and IGF-1 protein concentrations in the supernatant collected from mAstrocytes and ADSCs after separated co-culture were quantified by sandwich enzyme-linked immunosorbent assay (ELISA) using commercial ELISA kits.
For VEGF-A measurement, the mouse VEGF-A Platinum ELISA kit (BMS619/2, Thermo Fisher Scientific) was used according to the manufacturer’s instruction. IGF-I protein concentration was quantified with the mouse IGF-1 ELISA Kit (EMIGF1, Thermo Fisher Scientific) according to the manufacturer’s instructions. Before starting the protocol, samples stored at −80° were thawed on ice and diluted with the assay diluent provided in the kit. The standard and each sample were loaded in duplicate into the specific antibody-pre-coated 96-well strip plate provided in the kit and the manual instructions followed. At the end of the protocol, absorbance was read at 450 nm with the PHERAStar microplate reader (BMG, Labtech) and absorbance values transferred to an Excel data sheet. Data were finally analyzed with GraphPad 7. Briefly, the optical density value from the control (medium alone) was subtracted from the standard and sample readings. Data were then plotted on GraphPad 7, and a nonlinear sigmoidal standard curve (Curve Fit) was fitted. Protein concentration in the samples was then extrapolated and normalized to the number of cells counted at the end of the co-culture.