The Lipocalin family comprises a diverse group of mostly secreted soluble proteins that bind hydrophobic ligands and act as transporters,carrying small molecules to specific cells. Lipocalins are related by possessing an 8-stranded beta-barrel structure. Lipocalin-1,also named tear lipocalin (TL),von Ebners gland protein (VEG) and tear pre-albumin,binds a large number of hydrophobic molecules and exhibits cysteine proteinase inhibitor and endonuclear activities. Lipocalin-2,also known as neutrophil gelatinase-associated lipocalin (NGAL),is a component of granules in neutrophils from tissues that are normally exposed to microorganisms and is upregulated during inflammation. Lipocalin-2 can form homodimers and can heterodimerize with the neutrophil gelatinase MMP-9.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Human Lipocalin-2/NGAL BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human Lipocalin-2/NGAL concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:70pg/mlRange:156pg/ml-10000pg/mlSpecificity:Recognizes human Lipocalin-2/NGAL.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-h Lipocalin-2/NGAL monoclonal coating antibody is adsorbed onto microwells. Lipocalin-2/NGAL present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h Lipocalin-2/NGAL antibody is added and binds to Lipocalin-2/NGAL captured by the first antibody. Following incubation unbound Biotinylated anti-h Lipocalin-2/NGAL antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h Lipocalin-2/NGAL antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of Lipocalin-2/NGAL present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven Lipocalin-2/NGAL standard dilutions and Lipocalin-2/NGAL sample concentration determined.Kit Components:Aluminium pouches with a Microwell Plate: 8x12Standard: 2xVialBiotinylated antibody: 2xVialStreptavidin-HRP: 2xVialAssay Buffer (25ml/Vial): 2xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x12mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 3xstripsInstruction: 1xStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product,centrifuge the original vial after thawing and prior to removing the cap.