Human HNP1-3 (Neutrophil Peptide 1-3) ELISA Kit (HUES03176)

The Assay Genie sandwich ELISA quantitatively detects HNP1, HNP2, and HNP3 collectively as the HNP1-3 complex, with capture and detection antibodies pre-adsorbed on a strip-well microplate to minimize cross-reactivity between individual defensin isoforms. Standard curves are generated across a working range suitable for both baseline and elevated defensin concentrations, and the protocol recommends running each sample in duplicate alongside a freshly prepared seven-point standard series.

For accurate quantification, serum and plasma samples are typically diluted 1:100 to 1:1,000 in the supplied assay buffer depending on the expected analyte level, and spiking recovery experiments are advisable when working with less common matrices such as bronchoalveolar lavage fluid, wound exudate, or synovial fluid. The detection system uses a streptavidin-HRP conjugate with a TMB-based chromogenic substrate read at 450 nm after addition of an acidic stop solution, with intra-assay and inter-assay precision typically reported as coefficient of variation percentages on the lot-specific certificate of analysis.

HNP1-3 concentrations are commonly evaluated as biomarkers in research on bacterial and fungal infection, severe sepsis and septic shock, inflammatory bowel disease (notably ulcerative colitis), cystic fibrosis lung disease, acute respiratory distress syndrome, and chronic inflammatory disorders including rheumatoid arthritis and periodontitis. Because alpha-defensins are released rapidly from neutrophil azurophilic granules upon degranulation, circulating levels can rise within hours of an acute inflammatory stimulus, making the kit useful for kinetic studies of innate immune activation as well as longitudinal monitoring in clinical research cohorts.