Elisa kit against human bdnf

The amount of immobilized and released BDNF was quantified by an enzyme-linked immunosorbent assay (ELISA) kit against human BDNF (Boster Biological Technology Co., Ltd., Pleasanton, United States). The BDNF-ELISA kit was used in accordance to the manufacturer’s recommendations. A brief summary of the procedure follows. The 96-well plate delivered had been pre-coated with a monoclonal antibody for BDNF. Standards and samples were diluted in sample dilution buffer and added to the wells for 90 min. After the incubation, the plate content was discarded and a working solution of a biotinylated anti-human BDNF antibody was added for 60 min. After three washing steps with PBS, an incubation with a working solution containing an avidin-biotin-peroxidase complex followed (30 min). Further washing steps with PBS to remove the unbound complex were performed. For colorimetric detection 3,3′,5,5′-tetramethylbenzidine was added. The color development was stopped with 2 M sulfuric acid. All incubation steps were performed at 37°C. Absorbance was recorded at 450 nm on an EON spectrophotometer (Biotek, Winooski, United States).

For the quantification of the amount of immobilized and released BDNF an enzyme-linked immunosorbent assay (ELISA) kit against human BDNF (Boster Biological Technology Co., Ltd., Pleasanton, USA) was applied. The BDNF-ELISA kit was used in accordance to the manufacturer´s recommendations. In brief, a monoclonal antibody for BDNF had been precoated onto the 96-well plate. Standards and samples were diluted in sample dilution buffer and were added to the wells. After an incubation period of 90 min, the plate content was discarded and a working solution of a biotinylated anti-human BDNF antibody was added for 60 min. This was followed by three washing steps with PBS and by the incubation with a working solution containing an avidin-biotin-peroxidase complex. Then, the unbound conjugates were washed off with PBS. Finally, the detection was performed with 3,3´,5,5´-tetramethylbenzidine and stopped with 2 M sulphuric acid. All incubation steps were performed at 37°C. Absorbance was read at 450 nm on an EON spectrophotometer (Biotek, Winooski, USA).