Multiplex Assay Kit for Neutrophil gelatinase-associated lipocalin (NGAL) ,etc. by FLIA (Flow Luminescence Immunoassay)

Lipocalin-2; LCN2; p25; Lipocalin 2; Oncogene 24p3; 25 kDa alpha-2-microglobulin-related subunit of MMP-9; Siderocalin LCN2

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Neutrophil gelatinase-associated lipocalin (NGAL) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Neutrophil gelatinase-associated lipocalin (NGAL) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Neutrophil gelatinase-associated lipocalin (NGAL) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Neutrophil gelatinase-associated lipocalin (NGAL) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-98 93
EDTA plasma(n=5) 94-103 98
heparin plasma(n=5) 89-96 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Neutrophil gelatinase-associated lipocalin (NGAL) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Neutrophil gelatinase-associated lipocalin (NGAL) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Neutrophil gelatinase-associated lipocalin (NGAL) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 94-102% 93-103% 81-97% 87-101%
EDTA plasma(n=5) 98-105% 92-105% 82-97% 89-96%
heparin plasma(n=5) 80-101% 88-101% 91-99% 87-94%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:NGAL) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
International Journal of Nephrology and Renovascular Disease Correlation of serum neutrophil gelatinase-associated lipocalin with acute kidney injury in hypertensive disorders of pregnancy PubMed: PMC3794962
International Journal of Endocrinology Cholesterol Contributes to Diabetic Nephropathy through SCAP-SREBP-2 Pathway Pubmed: 24369464
International Journal of Sport Studies Lipocalin-2: Response To Resistance Training in Obese Young Men Ijssjournal: Source
Asian Journal of Sports Medicine Effects of Resistance versus Endurance Training on Plasma Lipocalin-2 in Young Men Tums: Source
European Respiratory Journal Differences in plasma and sputum biomarkers between COPD and COPD-asthma overlap Pubmed: 23794464
Acta Histochem.? Suppressor of cytokine signaling (SOCS) 2 attenuates renal lesions in rats with diabetic nephropathy Pubmed:24802166
Int Urol Nephrol. Nephrotoxic effects of varenicline as the most effective drug used for smoking cessation: a preliminary experimental study Pubmed:25812822
世界华人消化杂志 血清及胆汁中NGAL 对于区分良恶性胆道狭窄的临床意义 Wjgnet:Source
Diabetes &amp; Vascular Disease Research Urinary Kim-1 is a sensitive biomarker for the early stage of diabetic nephropathy in Otsuka Long-Evans Tokushima Fatty rats Sagepub:Source
Biomedical &amp; Pharmacology Journal Hs-CRP and Adipokin (Lcn2): Response to Exercise Training in Obese Men Biomedpharmajournal:Source
Asian Journal of Physical Education and Computer Science in Sports A Study On Thyroid Hormone And Its Effects On Exercise Ifcss:Source
临床儿科杂志 中性粒细胞明胶酶相关脂质运载蛋白、肾损伤分子1和白介素18 在诊断心肺分流术后急性肾损伤中的意义 Jcp-Sh:Source
Correlation of serum neutrophil gelatinase associated lipocalin with disease severity in hypertensive disorders of pregnancy Pubmed:Pmc4260273
Int J Chron Obstruct Pulmon Dis. Increased serum levels of lipocalin-1 and-2 in patients with stable chronic obstructive pulmonary disease Pubmed:Pmc4043430
Pediatric Anesthesia? Impact of 6% hydroxyethyl starch 130/0.42 and 4% gelatin on renal function in a pediatric animal model Pubmed:24916275
Inflammation.? Systemic and Flap Inflammatory Response Associates with Thrombosis in Flap Venous Crisis Pubmed:25448261
Int J Clin Exp Pathol Rosiglitazone attenuates renal injury caused by hyperlipidemic pancreatitis PubMed: 26191125
BMC Anesthesiol Increased cardiac index attenuates septic acute kidney injury: a prospective observational study PubMed: 25745357
中华临床医生杂志 中性粒细胞明胶酶相关脂质运载蛋白对腹膜透析相关性腹膜炎疗效监测的意义 Journal: Fulltext
PLoS One Ameliorative Effect of Chrysin on Adenine-Induced Chronic Kidney Disease in Rats PubMed: 25909514
Pediatric Anesthesia Impact of high doses of 6% hydroxyethyl starch 130/0.42 and 4% gelatin on renal function in a pediatric animal model Pubmed:26714833
Int Wound J. From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases. Pubmed:26991748
Archives of Biological Sciences Diabetes-induced renal failure is associated with tissue inflammation and neutrophil gelatinase-associated lipocalin: Effects of resveratrol publication:299492265_Diabetes-induced_renal_failu
International Journal of Molecular Medicine The neutrophil elastase inhibitor, sivelestat, attenuates sepsis-related kidney injury in rats pubmed:27430552
Hepatology. Hyperoxidized albumin modulates neutrophils to induce oxidative stress and inflammation in severe alcoholic hepatitis. pubmed:27775820
Lipids in Health and Disease Advanced glycation end products (AGEs) increase renal lipid accumulation: a pathogenic factor of diabetic nephropathy (DN) pubmed:28659153
International Wound Journal From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases pubmed:26991748
Clin Exp Pharmacol Physiol. Early urinary biomarkers of renal tubular damage by a high-salt intake independent of blood pressure in normotensive rats. pubmed:29027259
Scientific Reports Hemojuvelin Predicts Acute Kidney Injury and Poor Outcomes Following Cardiac Surgery Pubmed:29386545
Life Sciences Pyrrolidine dithiocarbamate inhibits mouse acute kidney injury induced by diclofenac by targeting oxidative damage, cytokines and NF-κB activity Pubmed:30036488
Molecules Effect of Pinocembrin Isolated from Mexican Brown Propolis on Diabetic Nephropathy Pubmed:29642511
American Journal of Translational Research Improved approach for normothermic machine perfusion of cold stored kidney grafts Pubmed:30018731
Chronic obstructive pulmonary disease (COPD)--from biomarkers to clinical phenotypes ISBN:978-951-51-4039-5
Disease Markers Potential Novel Biomarkers of Obstructive Nephropathy in Children with Hydronephrosis Doi: 10.1155/2018/1015726
Kidney and Blood Pressure Research The Kidney Specific Protein myo-Inositol Oxygenase, a Potential Biomarker for Diabetic Nephropathy Pubmed: 30504713
BMC Complementary and Alternative Medicine Dual actions on gout flare and acute kidney injury along with enhanced renal transporter activities by Yokuininto, a Kampo medicine Pubmed: 30871515
Naunyn-Schmiedeberg's archives of pharmacology Piceatannol protects against cisplatin nephrotoxicity via activation of Nrf2/HO-1 pathway and hindering NF-κB inflammatory cascade Pubmed: 31197431
Diabetes, Metabolic Syndrome and Obesity A small molecule inhibitor MCC950 ameliorates kidney injury in diabetic nephropathy by inhibiting NLRP3 inflammasome activation Pubmed: 31447572
Environment International Toxicity assessment due to prenatal and lactational exposure to lead, cadmium and mercury mixtures Pubmed: 31639605
Scientific Reports Alkaline phosphatase treatment of Acute Kidney injury in an infant piglet Model of cardiopulmonary Bypass with Deep Hypothermic circulatory Arrest Pubmed: 31578351
Int Urol Nephrol The evaluation of acute kidney injury due to ischemia by urinary neutrophil gelatinase-induced lipocalin (uNGAL) measurement in patients who underwent partial … Pubmed: 32980930
human & experimental toxicology Upregulated miR-101 inhibits acute kidney injury–chronic kidney disease transition by regulating epithelial–mesenchymal transition Pubmed: 32633566
INTERNATIONAL JOURNAL OF ENDOCRINOLOGY CОСТОЯНИЕ ТИРЕОИДНОЙ ФУНКЦИИ У ПАЦИЕНТОВ С САХАРНЫМ ДИАБЕТОМ 1-ГО ТИПА И ХРОНИЧЕСКОЙ БОЛЕЗНЬЮ ПОЧЕК …
INTERNATIONAL JOURNAL OF ENDOCRINOLOGY Thyroid function in patients with type 1 diabetes mellitus and chronic kidney disease receiving renal replacement therapy
Pediatr Int Early Kidney Injury in Immunoglobulin A Vasculitis: Role of Renal Biomarkers 33423326
ARCHIVES OF TOXICOLOGY Nrf2 deficiency aggravates the kidney injury induced by subacute cadmium exposure in mice 33398418
Biomed Res Int Renal Protective Effect of Beluga Lentil Pretreatment for Ischemia-Reperfusion Injury 33604384
International Journal of Medical Sciences Elevated plasma level of neutrophil gelatinase-associated lipocalin (NGAL) in patients with breast cancer 34104101
Intensive Care Medicine Experimental Preservation of renal endothelial integrity and reduction of renal edema by aprotinin does not preserve renal perfusion and function following experimental … 34169407
Biomolecules Preservation of Mitochondrial Coupling and Renal Function by Controlled Oxygenated Rewarming of Porcine Kidney Grafts 34944524
RSC Adv SERS based Y-shaped aptasensor for early diagnosis of acute kidney injury Pubmed:35733690
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