QuicKey Human NGAL(Neutrophil Gelatinase Associated Lipocalin) ELISA Kit

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This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human NGAL. Samples (or Standards) and biotinylated detection antibody specific for Human NGAL are added to the micro ELISA plate wells. Human NGAL would combine with the specific antibody. Then Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human NGAL, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. The OD value is proportional to the concentration of Human NGAL. You can calculate the concentration of Human NGAL in the samples by comparing the OD of the samples to the standard curve.

An unopened kit can be stored at 2-8℃ for six months. After test, the unused wells and reagents should be stored according to the table below.

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human NGAL were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NGAL were tested on 3 different plates, 20 replicates in each plate.

The recovery of Human NGAL spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Neutrophil gelatinase associated lipocalin (NGAL) also known as Lipocalin 2 or Lcn2 is a 25 kDa, is a member of the lipocalin superfamily and a pleiotropic mediator of various inflammatory processes [1].NGAL, identified originally as a protein associated with matrix metalloproteinase 9 (MMP-9) of human neutrophils. Lipocalins are extracellular proteins which share a common tertiary structure that forms a barrel-like hydrophobic ligand binding site. When bound to MMP-9, NGAL protects it from proteolytic degradation sustaining the proteolytic activity of MMP-9. Neutrophil gelatinase-associated lipocalin (NGAL), is released by various cell types and is an attractive biomarker of inflammation, ischemia, infection, and kidney damage. Both intestinal and metabolic inflammation, as observed in obesity and related disorders, are associated with increased NGAL synthesis [2]. While NGAL in the intestinal tract regulates the composition of the gut microbiota and shows anti-inflammatory activities, it also exhibits proinflammatory activities in other experimental settings. Plasma NGAL is a novel marker of kidney function, which correlates to duration of end-stage renal failure (ESRD) and serum creatinine in uremic patients awaiting kidney transplantation. Plasma NGAL is associated with homocysteine in transplanted patients.

1. Kjeldsen L, Cowland J B, Borregaard N. Human neutrophil gelatinase-associated lipocalin and homologous proteins in rat and mouse.[J]. Biochim Biophys Acta, 2000, 1482(1):272-283.

2. Tilg, H., & Moschen, A. R. Adipocytokines: mediators linking adipose tissue, inflammation and immunity. Nature Reviews Immunology, 2006, 6(10), 772.