Lipocalin-2/NGAL Human ELISA

Regulatery Status: RUO
Type: Sandwich ELISA, Biotin-labelled antibody
Other Names Status: 24p3, MSFI, LCN2
Species: Human
Catalog No Size
Product Catalog No: RD191102200R Pack Size: 96 wells (1 kit)

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Product Features

Lipocalin-2 (LCN2) is a 25 kDa secretory glycoprotein, also called NGAL (neutrophil gelatinase-associated lipocalin); NL (neutrophil lipocalin); p25; oncogen 24p3 and 25 kDa alpha-2-microglobulin-related subunit of MMP-9 (LCN2 forms a covalently linked, disulfide-bridged heterodimer with the 92 kDa type V collagenase (MMP-9)). LCN2 is predominantly expressed in adipose tissue and liver. It belongs to the lipocalin superfamily that consists of over 20 small secretory proteins. Lipocalin folds consist of 8 antiparallel ß-sheets that surround a hydrophobic pocket. A common feature of this protein family, following from its structure, is its capacity to bind and transport small lipophilic substancies such as free fatty acids, retinoids, arachidonic acid and various steroids. Although Lipocalin-2 was identified more than a decade ago, the physiologic function of this protein remains poorly understood. LCN2 appears to be upregulated in cells under the “stress” (e.g. from infection, inflammation, in tissues undergoing involution to ischemia or neoplastic transformation). Plasma levels of LCN2 rise in inflammatory or infective condition. It mediates an immune response to bacterial infection by sequestering iron. In this case, LCN2 may represent a promising candidate as a therapeutic agent against bacterial infection. Several recent reports suggest that LCN2 might represent a sensitive biomarker for early renal injury. In cardiopulmonary bypass-induced acute renal injury and cisplatin-induced nephrotoxic injury, increased de novo synthesis of LCN2 in proximal tubule cells leads to sharply increased concentration of this protein in both urine and serum. LCN2 might also be critical for normal kidney formation in the earliest stages of mammalian development. LCN2 may play an important role in breast cancer, in complex with MMP-9, by protecting MMP-9 from degradation thereby enhancing its enzymatic activity and facilitating angiogenesis and tumor growth. LCN2 is also highly expressed after malignant transformation of the lung, colon and pancreatic epithelia. Circulating levels of LCN2 play a causative role in pathogenesis of obesity-induced metabolic disorders such as insulin resistance, Type 2 Diabetes Mellitus and cardiovascular disorders. In addition, serum LCN2 concentrations were positively associated with adipocyte-fatty acid binding protein (A-FABP), a novel serum marker for adiposity and metabolic syndrome.

Technical Sheet / Info

Intended use

The RD191102200R Human Lipocalin-2/NGAL ELISA is a sandwich enzyme immunoassay for the quantitative measurement of human lipocalin-2 in serum, plasma and urine samples.

  • The total assay time is less than 3.5 hours
  • The kit measures lipocalin-2 in serum, plasma (EDTA), urine samples and stool extract
  • Assay format is 96 wells
  • Quality Controls are human serum based. No animal sera are used
  • Standard is recombinant protein based
  • Components of the kit are provided ready to use, concentrated or lyophilized

Clinical Application

  • Renal injury
  • Angiogenesis
  • Oncology
  • Diabetes mellitus
  • Metabolic syndrome

Test principle

In the BioVendor Human Lipocalin-2/NGAL ELISA, standards, quality controls and samples are incubated in microplate wells pre-coated with polyclonal anti-human lipocalin-2 antibody. After one hour incubation and washing, biotin labelled polyclonal anti-human lipocalin-2 antibody is added and incubated with captured lipocalin-2 for one hour. After another washing, streptavidin-HRP conjugate is added. After 30 minutes incubation and the last washing step, the remaining conjugate is allowed to react with the substrate solution (TMB). The reaction is stopped by addition of acidic solution and absorbance of the resulting yellow product is measured. The absorbance is proportional to the concentration of lipocalin-2. A standard curve is constructed by plotting absorbance values against concentrations of standards, and concentrations of unknown samples are determined using this standard curve.

Summary of protocol

  • Reconstitute QCs and Master Standard and prepare set of Standards
  • Dilute samples 30×
  • Add 100 μl Standards, QCs and samples
  • Incubate at RT for 1 hour/300 rpm
  • Wash plate 3 times
  • Add 100 μl Biotin Labelled Antibody
  • Incubate at RT for 1 hour/300 rpm
  • Wash plate 3 times
  • Add 100 μl Streptavidin HRP Conjugate
  • Incubate at RT for 30 minutes/300 rpm
  • Wash plate 3 times
  • Add 100 μl Substrate Solution
  • Incubate at RT for 10 min
  • Add 100 μl Stop Solution
  • Read absorbance and calculate results

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    References

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    – Borkham-Kamphorst E, van de Leur E, Zimmermann HW, Karlmark KR, Tihaa L, Haas U, Tacke F, Berger T, Mak TW, Weiskirchen R. Protective effects of lipocalin-2 (LCN2) in acute liver injury suggest a novel function in liver homeostasis. Biochim Biophys Acta. 2013 May;1832 (5):660-73

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    – Yavuz S, Anarat A, Acarturk S, Dalay AC, Kesiktas E, Yavuz M, Acarturk TO. Neutrophil gelatinase associated lipocalin as an indicator of acute kidney injury and inflammation in burned children. Burns. 2014 Jun;40 (4):648-54

    – Zorlu M, Kiskac M, Karatoprak C, Kesgin S, Cakirca M, Yildiz K, Ardic C, Cikrikcioglu MA, Erkoc R. Assessment of serum apelin and lipocalin-2 levels in patients with subclinical hypothyroidism. Ann Endocrinol (Paris). 2014 Feb;75 (1):10-4

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