QuicKey Human NGAL(Neutrophil Gelatinase Associated Lipocalin) ELISA Kit

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Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	2.5 h
Reactivity	Human
Detection method	Colormetric
Colormetric	0.13-8 ng/mL
Sensitivity	0.06 ng/mL
Sample volume	50μL/well
Sample type	serum, plasma, urine
Specificity	This kit recognizes Human NGAL in samples. No significant cross-reactivity or interference between Human NGAL and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	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.

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. (The specific components might be slightly different in delivered kits, please refer to the manual for more information.)

Item	Specifications	Storage conditions after test
Micro ELISA Plate (Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	2-8℃, 1 month
Reference Standard	96T: 2 vials 48T: 1 vial	Discard unused reconstituted standard and dilutions
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8℃
Biotinylated Detection Ab Working Solution	1 vial, 6 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 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).

1. Microplate reader with 450nm wavelength filter

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

3. Incubator capable of maintaining 37℃

4. Deionized or distilled water

5. Absorbent paper

6. Loading slot for Wash Buffer

Concentration(ng/mL)	8	4	2	1	0.5	0.25	0.13	0
OD	2.569	2.13	1.533	1.088	0.656	0.437	0.225	0.059
Corrected OD	2.51	2.071	1.474	1.029	0.597	0.378	0.166	-

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.

Immunology, Cancer, Signaling transduction

SwissProt	P80188 Human
Entrez Gene	3934 Human

LCN2, Lipocalin 2, Oncogene 24p3, MSFI