The Biosensis Mouse and Rat proNGF Rapid enzyme-linked immunosorbent assay (ELISA) Kit is a sandwich ELISA that allows the quantification of rodent full-length proNGF protein in less than 4 hours in cell culture supernatants, cell lysates and tissue homogenates only if used as directed. This ELISA Kit has also successfully been used on mouse urine samples (Ryu JC et al., 2018). Please refer to the kit protocol for specific use instructions for each substrate application.

This ELISA kit contains a recombinant mouse proNGF standard expressed in E.coli and consists of a pre-coated anti-proNGF capture antibody, a biotinylated anti-proNGF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The addition of a substrate (3,3',5,5'-tetramethylbenzidine, TMB) yields a colored reaction product which is directly proportional to the concentration of proNGF present in samples and protein standards. A proNGF positive control (QC sample) is provided to assure consistent assay performance. This ELISA kit detects rat proNGF due to high degree of homology (96%) with mouse proNGF based on amino acid sequence, and the ability of this kit in detecting proNGF in rat PC12 cell lysates and rat brain tissue homogenate. In the absence of a true rat proNGF standard, results may be expressed as 'mouse proNGF equivalents'. This ELISA kit shows only 20% reactivity with the human form of proNGF and is therefore not suitable to quantify human proNGF. No cross-reactivity was observed with mature mouse NGF and full-length proBDNF when tested in assay buffer.

The antibodies used in this ELISA kit bind epitopes within the pro-domain (capture) and mature domain (detection) of the protein, thus this ELISA assay does not detect the pro-domain peptide. This kit has not been tested for other applications. Sufficient amount of proNGF standard is supplied to allow for spike- and recovery experiments in order to validate this ELISA assay for other sample matrices if required.

Please refer to the kit protocol for specific use instructions for each substrate application.