Rat neutrophil gelatinase-associated lipocalin,NGAL ELISA Kit

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Name: Rat neutrophil gelatinase-associated lipocalin,NGAL ELISA Kit
Brand: CUSABIO
SKU: CSB-E09409r

Product Details

Description

The Rat neutrophil gelatinase-associated lipocalin (NGAL) ELISA Kit is used to quantitatively measure NGAL concentrations in rat serum, plasma, urine, or tissue homogenates. It performs well in important characteristics, including sensitivity, specificity, precision, recovery, linearity, and lot-to-lot consistency. This assay is based on the sandwich ELISA mechanism and enzyme-substrate chromogenic reaction. The solution color develops proportionally to the amount of NGAL in the sample. And the intensity of the color can be measured at 450 nm via a microplate reader.

NGAL was originally isolated from secondary granules of human neutrophils as a 25 KDa protein covalently linked to matrix metalloproteinase-9 (MMP-9) in human neutrophils. It is highly induced in the kidney after ischemic or nephrotoxic acute kidney injury (AKI) in animal models. NGAL has been reported as an early urine and plasma biomarker of AKI in pediatric and adult cardiac surgery. Urinary NGAL has been employed as a routine measurement in AKI patients by King Chulalongkorn Memorial Hospital since 2015.

Target Name

lipocalin 2

Alternative Names

Lcn2 ELISA Kit; Neutrophil gelatinase-associated lipocalin ELISA Kit; NGAL ELISA Kit; Alpha-2-microglobulin-related protein ELISA Kit; Alpha-2U globulin-related protein ELISA Kit; Lipocalin 24p3 ELISA Kit; Lipocalin-2 ELISA Kit; Siderocalin LCN2 ELISA Kit; p25 ELISA Kit

Abbreviation

LCN2

Uniprot No.

P30152

Species

Rattus norvegicus (Rat)

Sample Types

serum, plasma, urine, tissue homogenates

Detection Range

0.312 ng/mL-20 ng/mL

Sensitivity

0.078 ng/mL

Assay Time

1-5h

Sample Volume

50-100ul

Detection Wavelength

450 nm

Research Area

Immunology

Assay Principle

quantitative

Measurement

Sandwich

Precision

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of rat NGAL in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
	Sample	Serum(n=4)
1:1	Average %	88
	Range %	80-98
1:2	Average %	95
	Range %	89-99
1:4	Average %	85
	Range %	81-90
1:8	Average %	104
	Range %	99-107

Recovery

The recovery of rat NGAL spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.

Sample Type	Average % Recovery	Range
Serum (n=5)	98	93-104
EDTA plasma (n=4)	95	90-100

Typical Data

These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

ng/ml	OD1	OD2	Average	Corrected
20	2.451	2.498	2.475	2.287
10	1.954	1.967	1.961	1.773
5	1.226	1.269	1.248	1.060
2.5	0.758	0.789	0.774	0.586
1.25	0.529	0.505	0.517	0.329
0.625	0.380	0.351	0.366	0.178
0.312	0.279	0.271	0.275	0.087
0	0.187	0.189	0.188

Materials provided

A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-rat NGAL. This dismountable microplate can be divided into 12 x 8 strip plates.
Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
One vial Biotin-labeled NGAL antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
One vial Sample Diluent(50 ml/bottle)---Dilute the sample to an appropriate concentration.
One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
An instruction manual

Materials not provided

A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
An incubator can provide stable incubation conditions up to 37°C±5°C.
Centrifuge
Vortex
Squirt bottle, manifold dispenser, or automated microplate washer
Absorbent paper for blotting the microtiter plate
50-300ul multi-channel micropipette
Pipette tips
Single-channel micropipette with different ranges
100ml and 500ml graduated cylinders
Deionized or distilled water
Timer
Test tubes for dilution

Troubleshooting
and FAQs

ELISA kit FAQs

Storage

Store at 2-8°C. Please refer to protocol.

Lead Time

3-5 working days

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Target Data

Function	Iron-trafficking protein involved in multiple processes such as apoptosis, innate immunity and renal development. Binds iron through association with 2,5-dihydroxybenzoic acid (2,5-DHBA), a siderophore that shares structural similarities with bacterial enterobactin, and delivers or removes iron from the cell, depending on the context. Iron-bound form (holo-24p3) is internalized following binding to the SLC22A17 (24p3R) receptor, leading to release of iron and subsequent increase of intracellular iron concentration. In contrast, association of the iron-free form (apo-24p3) with the SLC22A17 (24p3R) receptor is followed by association with an intracellular siderophore, iron chelation and iron transfer to the extracellular medium, thereby reducing intracellular iron concentration. Involved in apoptosis due to interleukin-3 (IL3) deprivation
Gene References into Functions	NGAL might be related to attenuated tubular epithelial cell damage via adjusting autophagy. PMID: 29367586 The results showed that remote limb preconditioning could decrease the expression of LCN2 protein, but having no obvious effects on its receptor except the time point of 72h after cerebral ischemia. PMID: 29122651 High LCN2 expression is associated with stress-induced mitochondrial dysfunction and heart failure. PMID: 29378951 This study is to investigate the level of Neutrophil gelatinase-associated lipocalin (NGAL) and autophagy status during renal Ischemia/reperfusion injury. PMID: 29704511 This study demonstrated that Neutrophil gelatinase-associated lipocalin and microglial activity are associated with distinct postoperative behavioral changes in rats. PMID: 27865916 LCN-2 protein levels were markedly increased in the ipsilateral cortex and hippocampus after traumatic brain injury. Most LCN-2-positive cells appeared to be astrocytes. PMID: 26463963 Nitric oxide level affects LCN-2 expression via regulation of LCN-2 protein stability under inflammatory condition and LCN-2 may reduce beta-cell viability by promoting apoptosis. PMID: 27233602 The NGAL molecule showed upregulation in the degeneration process after nerve injury, so it may play an important role in nerve repair PMID: 27087673 The LCN2 may be a potentially useful biomarker for lung hypoplasia in a rat and human Congenital diaphragmatic hernia (CDH). PMID: 27592368 Hypertension and hyperglycemia synergize to cause incipient renal tubular alterations resulting in increased NGAL urinary excretion.NGAL urinary excretion is also increased in hypertensive-diabetic rats. PMID: 25148248 This is the first report on Pla2g4c gene-deficient rats and their low susceptibility to mammary carcinogenesis by enhancing NF-kappaB/Lcn2-induced apoptosis. PMID: 26013918 Epidural anesthesia was associated with an increase in the expression lipocalin-2 and a decrease in the expression of E coli DNA at pseudosurgical sites in sick but not healthy rats. PMID: 25782996 Repeated administration of gentamicin to male SD rats for 1, 3, or 7 days resulted in a dose- and time-dependent increase in the expression of KIM-1 and NGAL. PMID: 25087119 dietary fructose leads to an overexpression of lipocalin-2 in rat fatty liver PMID: 24587658 Lcn2 is an important regulator in small-for-size fatty liver graft injury and targeting Lcn2 may be feasible for preventing marginal graft failure in LDLT. PMID: 24374540 NGF-treated tissue showed increased expression of LCN2 which was inhibited by the presence of Ro 08-2750 in intervertebral disk cells. PMID: 25062286 lipocalin-2 is released by injured neurons as a help me distress signal that activates microglia and astrocytes into potentially prorecovery phenotypes. PMID: 24916903 r24p3-R/hNGAL-R may represent a high-affinity multi-ligand receptor for apical internalization and transcytosis of intact proteins/peptides by the lower intestine. PMID: 23940770 Urinary KIM-1, NGAL and NAG were compared against serum creatinine and renal histological score in the initiation, maintenance, and recovery phases of cisplatin (CP)-induced acute kidney injury. PMID: 23360846 We studied Lcn2 gene expression during liver regeneration after partial hepatectomy(PH). Lcn2 gene expression showed upward curves from 4 to 72 h after PH in both 70% and 40% PH rats and peaked at 12 h (8 times vs 0 h). PMID: 23481292 The expression of NGAL significantly increases in renal interstitial fibrosis, and is closely correlated with MMP-9, TNF-alpha and TGF-beta1. PMID: 23158800 lcn2 is induced in activated astrocytes in response to neurodegeneration, but its induction is independent of TDP-43 or FUS expression in astrocytes PMID: 23431168 High NGAL is associated with proteinuria. PMID: 21551085 Acute myocardial infarct in the chronic kidney disease setting is associated with accelerated renal fibrosis and long-term elevated urine Ngal values. PMID: 22889806 Exposure of Wistar rats to methyl tertiary-butyl ether in drinking water resulted in limited renal changes in male rats suggestive of alpha(2u)-globulin nephropathy. PMID: 22833177 Adoptive transfer of IL-10 macrophages to ischemic kidneys blunts acute kidney injury through induction of lipocalin-2. PMID: 22278021 LIPOCALIN-2 is a major acute-phase protein in a rat and mouse model of sterile abscess PMID: 22249220 Lcn2-induced cardiomyocyte apoptosis is of physiological significance and occurs via a mechanism involving elevated intracellular iron levels and Bax translocation. PMID: 22117066 An evaluation of the SIP24 and MPO distribution and levels of expression in the rat uterus is consistent with the hypothesis that SIP24 modulates the neutrophil population by activities such as inducing their apoptosis. PMID: 16546827 SMC-produced NGAL interacts with MMP-9, a mechanism by which NGAL may modulate MMP-9 proteolytic activity in the vascular repair process.[Neutrophil gelatinase-associated lipocalin, NGAL] PMID: 17148685 LCN2 expression shows clear correlation to liver damage and resulting inflammatory responses, rather than to the degree of liver fibrosis, which in fact may imply a distinct diagnostic value as an early biomarker of liver inflammation. PMID: 21457438 Cardiomyocytes, vascular wall cells and fibroblasts in myocarditis strongly express Lcn2/NGAL via proinflammatory cytokines PMID: 20057160 Acute phase response to acute endotoxemia involves induction of Lcn2 in both the lung and liver. PMID: 17490638 Show More Hide All
Subcellular Location	Secreted
Protein Families	Calycin superfamily, Lipocalin family
Database Links	KEGG: rno:170496 STRING: 10116.ENSRNOP00000018776 UniGene: Rn.11303

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Intra-assay Precision (Precision within an assay): CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess.

Inter-assay Precision (Precision between assays): CV%<10%

Three samples of known concentration were tested in twenty assays to assess.

Code	CSB-E09409r
Size	96T,5×96T,10×96T
Trial Size
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