Gdnf elisa kit

After 5 days of treatment with the indicated PRP, the culture medium was collected, and the concentrations of NGF-β, VEGF, and GDNF in the culture medium were determined using an NGF-β ELISA kit (Boster, EK0471), a VEGF ELISA kit (Boster, EK0540) and a GDNF ELISA kit (EK0363) as instructed by the manufacturer.

The medium for measuring the GDNF levels was collected after culturing goat SSCs for 48 h. The medium was frozen at −80 °C until analysis. GDNF levels were determined by using a Human glial cell line-derived neurotrophic factor (GDNF) ELISA Kit (BOSTER). Briefly, 100 μl of standard and sample were added per well in 96-well plates (12 × 8 coated Microwell) and incubated for 90 min at 37°C. Removed the liquid of each well and added 100 μl of Biotin-antibody to each well, and incubated for 90 min at 37°C. Each well was aspirated and washed 3 times with washing Buffer. ABC Buffer was added and incubated for 30 min at 37°C and washed 5 times. Finally, TMB was added and the GDNF levels were determined by absorbance at 450 nm. The standard curve indicated a direct relationship between optical density and GDNF concentration.

The collected culture mediums at 1, 3, 5, 7, and 10 days were used for determining the concentration of the neurotrophic factors secreted by live nerve microtissues. The concentrations of nerve growth factor-β (NGF-β), vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) were determined using an NGF-β ELISA kit (Boster, EK0471), a VEGF ELISA kit (Boster, EK0540), a BDNF ELISA kit (Boster, 0308) and a GDNF ELISA kit (Boster, EK0363) as instructed by the manufacturer.