预期应用
ELISA法定量测定人血清、血浆、细胞培养上清或其它相关生物液体中中性粒细胞防御素1-3(HNP1-3)含量。
实验原理
用纯化的抗体包被微孔板，制成固相载体，往包被抗HNP1-3抗体的微孔中依次加入标本或标准品、生物素化的抗HNP1-3抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的HNP1-3呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。 Intended use
This immunoassay kit allows for the use in vitro quantitative determination of human neutrophil peptide 1-3, HNP1-3 concentrations in cell culture supernates, serum, plasma and other biological fluids.
Introduction
Defensins are cytotoxic peptides and have a well-defined role in host defense. Human alpha defensins 1-3 (known as human neutrophil peptides (HNPs) 1-3) are primarily produced by peripheral neutrophils and constitute about 50% of the azurphil granule protein. Studies have suggested that peripheral neutrophils and the resident neutrophils in the ovary enhance the release of IL-8 and TNF-alpha that play a role in ovulation and influence fertilisation rate and IVF outcome.
Test principle
The microtiter plate provided in this kit has been pre-coated with an antibody specific to HNP1-3. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for HNP1-3 and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain HNP1-3, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of HNP1-3 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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