Description
Description: This assay employs a two-site sandwich ELISA to quantitate NPPB in samples. An antibody specific for NPPB has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any NPPB present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for NPPB is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of NPPB bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: NPPB is expressed in a subset of TRPV1 (602076)-expressing neurons and found that Nppb-null mice selectively lose almost all behavioral responses to itch-inducing agents. Nppb triggered potent scratching when injected intrathecally in wildtype and Nppb-null mice, showing that this neuropeptide evokes itch when released from somatosensory neurons. Itch responses were blocked by toxin-mediated ablation of Nppb receptor-expressing cells, but a second neuropeptide, gastrin-releasing peptide (GRP; 137260), still induced strong responses in the toxin-treated animals. Mishra and Hoon (2013) concluded that their results defined the primary pruriceptive neurons, characterized Nppb as an itch-selective neuropeptide, and revealed the next 2 stages of this dedicated neuronal pathway