QuicKey Human NGAL(Neutrophil Gelatinase Associated Lipocalin) ELISA Kit

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      Catalog number:E-TSEL-H0003

      Size:
      • 96T
      Qty:
      - +
      Price: $390

      Reactivity: Human

      Detection Range: 0.13~8 ng/mL

      Sensitivity: 0.06 ng/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

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      QuicKey Series

      Get more sensitive and precise results with saving at least 1h comparing to traditional ELISA Kits. The new developed technology in house will help to accelerate your science research in a more efficient way.


      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human NGAL concentrations in serum, plasma, urine. Please consult technical support for the applicability if other biological fluids need to be tested.



      Background

      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.


      Test principle

      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 combined 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.

      Assay type

      Sandwich-ELISA

      Format

      96T

      Assay time

      2.5h

      Reactivity

      Human

      Detection method

      Colormetric

      Detection range

      0.13-8 ng/mL

      Sensitivity

      0.06 ng/mL

      Sample volume

      100μL

      Sample type

      Serum, Plasma, Urine


      Specificity

      This kit recognizes Human NGAL in samples. No significant cross-reactivity or interference between NGAL and analogues was observed.


      Typical data

      As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.


      Concentration(ng/mL)

      8

      4

      2

      1

      0.5

      0.25

      0.125

      0

      OD

      2.494

      2.002

      1.353

      0.885

      0.559

      0.327

      0.192

      0.063

      Corrected OD

      2.431

      1.939

      1.290

      0.822

      0.496

      0.264

      0.129

      -



      Sample values

      Serum/Plasma/Urine–Samples from apparently healthy volunteers were evaluated for the presence of Human NGAL in this assay.

      Sample Type

      Source

      Range

      Dilution Factor

      Serum(n=19)

      Healthy human

      68.34-230.5ng/mL

      50-500

      EDTA plasma (n=12)

      Healthy human

      52-165.6ng/mL

      50-500

      Urine(n=17)

      Healthy human

      0.66-74.98ng/mL

      1-50


      Precision

      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.

       

      Intra-assay Precision

      Inter-assay Precision

      Sample

      1

      2

      3

      1

      2

      3

      n

      20

      20

      20

      20

      20

      20

      Mean(ng/mL)

      0.37

      0.96

      3.96

      0.33

      0.99

      3.63

      Standard deviation

      0.02

      0.05

      0.19

      0.02

      0.05

      0.16

      CV (%)

      5.41

      5.21

      4.8

      6.06

      5.05

      4.41



      Recovery

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

      Sample Type

      Range (%)

      Average Recovery (%)

      Serum (n=19)

      87-108

      97

      EDTA plasma (n=12)

      80-115

      90

      Urine(n=17)

      81-120

      97


      Linearity

      Samples were spiked with high concentrations of Human NGAL and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

       

       

      Serum (n=4)

      EDTA  plasma (n=4)

      Urine(n=4)

      1:2

      Range (%)

      105-108

      100-115

      91-108

      Average (%)

      106

      105

      100

      1:4

      Range (%)

      95-105

      97-117

      98-105

      Average (%)

      101

      104

      102

      1:8

      Range (%)

      96-105

      95-112

      84-105

      Average (%)

      101

      103

      93

      1:16

      Range (%)

      104-108

      94-119

      85-105

      Average (%)

      106

      107

      97


      Kit components & Storage

      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.

      Item

      Specifications

      Storage conditions after test

      Micro ELISA Plate (Dismountable)

      8 wells ×12 strips

      2-8, 1 months

      Reference Standard

      2 vials

      Discard unused reconstituted standard and dilutions

      Reference Standard & Sample Diluent

      1 vial, 20 mL

      2-8

      Biotinylated Detection Ab Working Solution

      1 vial, 12 mL

      HRP Conjugate Diluent

      1 vial, 14 mL

      Concentrated Wash Buffer (25×)

      1 vial, 30 mL

      Concentrated HRP Conjugate (100×)

      1 vial, 120 μL

      2-8(Protect from light)

      Substrate Reagent

      1 vial, 10 mL

      Stop Solution

      1 vial, 10 mL

      2-8

      Plate Sealer

      5 pieces

       

      Product Description

      1 copy

      Certificate of Analysis

      1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.

      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).


      Other supplies required

      Microplate reader with 450 nm wavelength filter

      High-precision transfer pipette, EP tubes and disposable pipette tips

      37℃ Incubator

      Deionized or distilled water

      Absorbent paper

      Loading slot for Wash Buffer


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