Description
Principle of the assay: Rat NGAL ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for NGAL has been precoated onto 96-well plates. Standards(NSO, Q21-N198) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for NGAL is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the rat NGAL amount of sample captured in plate.
Background: Lipocalin-2 (LCN2), also known as NGAL, is a protein associated with neutrophil gelatinase.1 The LCN2 gene islocated at 9q34 and contains 7 exons.2 The 25-kD LCN2 protein is believed to bind small lipophilic substances suchas bacteria-derived lipopolysaccharide (LPS) and formylpeptides and may function as a modulator of inflammation. NGAL tightly binds bacterial catecholate-type ferric siderophores through a cyclically permuted, hybrid electrostatic/cation-pi interaction and is a potent bacteriostatic agent in iron-limiting conditions.3 The primary LCN2transcript is 3,696 nucleotides long, and the processed transcript is 809 nucleotides long.4 LCN2 expression in adultbone marrow, uterus, prostate, salivary gland, stomach, appendix, colon, trachea, and lung, and in fetal spleen and lung. The standard product used in this kit is recombinant rat NGAL, consisting of 177 amino acids with the molecular mass of 21kDa