The Biosensis NGF Rapid™ enzyme-linked immunosorbent assay (ELISA) kit is a sandwich ELISA that allows the quantification of human NGF in less than 3 hours in cell culture supernatants, serum, plasma (citrate) and brain extracts only if used as directed.

This NGF Rapid™ ELISA Kit is designed to measure human NGF and thus employs a recombinant human NGF standard approved by the World Health Organization (WHO). Due to a high degree of NGF sequence homology, the antibodies used in this kit will also detect NGF from other species including mouse and rat.

The purpose of this kit is the in vitro quantitative determination of NGF in samples such as serum, plasma, cell culture supernatants and brain extracts.

The antibodies used in this ELISA kit bind epitopes within the mature domain of the protein and therefore can recognize both the pro- and the mature form of NGF. However, internal validation data demonstrates <0.1% cross-reactivity (determined at 25 ng/ml, diluted in assay buffer) with human proNGF protein, suggesting the preferential quantification of mature NGF over full-length human proNGF. Note that accurate NGF quantification in human citrate-plasma requires the addition of Heterophilic Antibody Blocker provided in the kit, and available for purchase separately.

This kit consists of a pre-coated mouse monoclonal anti-NGF capture antibody, a biotinylated anti-NGF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The addition of a substrate (3,3’,5,5’-tetramethylbenzidine, TMB) yields a coloured reaction product which is directly proportional to the concentration of NGF present in samples and protein standards. A human NGF positive control (QC sample) is provided to assure consistent assay performance.

Please refer to the kit protocol for specific use instructions for each substrate application, in particular serum and plasma samples.