ELISA for Monitoring Nerve Growth Factor

Nerve growth factor (NGF), a neurotrophin critical to neuronal viability, has become a popular research focus for the treatment of neurodegenerative diseases. Enzyme-linked immunosorbent assays (ELISA) allow the quantification of cellular proteins, such as NGF, secreted into conditioned culture media. However, off-the-shelf reagents and kit components may require optimization depending on the specific specimens and/or antibodies that will be utilized with the ELISA assay. Herein we describe a protocol for developing a sandwich ELISA, incorporating NGF-specific primary and secondary antibodies capable of detecting NGF at concentrations as low as 10 pg/mL.