ELISA Kit for Renalase (RNLS)

MAO-C; C10orf59; FAD-Dependent Amine Oxidase; Monoamine oxidase-C; alpha-NAD(P)H oxidase/anomerase

Specificity

This assay has high sensitivity and excellent specificity for detection of Renalase (RNLS).
No significant cross-reactivity or interference between Renalase (RNLS) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Renalase (RNLS) and the recovery rates were calculated by comparing the measured value to the expected amount of Renalase (RNLS) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-104 94
EDTA plasma(n=5) 89-103 93
heparin plasma(n=5) 85-101 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Renalase (RNLS) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Renalase (RNLS) 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 Renalase (RNLS) 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) 86-94% 80-97% 79-98% 97-105%
EDTA plasma(n=5) 88-99% 94-101% 84-102% 86-95%
heparin plasma(n=5) 84-98% 93-103% 90-97% 78-104%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Pol Arch Med Wewn. Renalase, kidney function, and markers of endothelial dysfunction in renal transplant recipients Cartelcolor: source
Angiology Renalase, Stroke, and Hypertension in Hemodialyzed Patients PubMed: 22583169
Peritoneal dialysis international Renalase in peritoneal dialysis patients is not related to blood pressure, but to dialysis vintage. Pdiconnect: Source
Archives of Medical Science Vascular adhesion protein-1 and renalase in regard to diabetes in hemodialysis patients PubMed: PMC3542495
Renal Failure Circulating Levels of Renalase, Norepinephrine, and Dopamine in Dialysis Patients Pubmed: 23530612
Borgis - Postepy Nauk Medycznych VAP-1 in chronic kidney disease and diabetic kidney disease Czytelniamedyczna: Source
Journal of Basic and Clinical Physiology and Pharmacology The relationship between dopamine and renalase in type 2 diabetic patients with and without diabetic nephropathy Pubmed: 15365253
Exp Biol Med (Maywood). Plasma and urine renalase levels and activity during the recovery of renal function in kidney transplant recipients. Pubmed: 24599883
Kidney Blood Press Res. Renalase, a novel enzyme involved in blood pressure regulation, is related to kidney function but not to blood pressure in hemodialysis patients. Pubmed: 22539018
Nephrology Dialysis Transplantation Renalase in hypertension and kidney disease Oxfordjournals: Source
Lipids in Health and Disease Markers of increased cardiovascular risk in patients with chronic kidney disease Biomedcentral:Source
Research &amp; Experimental? Plasma and urine renalase levels and activity during the recovery of renal function in kidney transplant recipients Pubmed:24599883
Pediatr Nephrol Normal values for urine renalase excretion in children Pubmed:25060760
J Am Soc Hypertens. Low plasma renalase concentration in hypertensive patients after surgical repair of coarctation of aorta Pubmed:25064768
Cardiology. Effect of salt intake and potassium supplementation on urinary renalase and serum dopamine levels in Chinese adults Pubmed:25824645
J Am Soc Hypertens Circulating renalase, catecholamines, and vascular adhesion PubMed: 26403854
Indian Pediatr Renalase levels in children with solitary functioning kidney PubMed: 26713989
Oxford Journals SP321 RENALASE, RENAL FUNCTION AND BIOMARKERS OF ENDOTHELIAL DYSFUNCTION IN CHRONIC KIDNEY DISEASE PATIENTS Content: 30
journal of hypertension PP.27.07: PREHYPERTENSIVE ALTERATIONS OF ARTERIAL AND VENOUS REACTIVITY IN A RAT POLYCYSTIC OVARY SYNDROME MODEL. EFFECTS OF PARALLEL VITAMIN D3 ADMINISTRATION Jhypertension: Abstract
journal of hypertension PP.27.06: RENALASE, RENAL FUNCTION AND BIOMARKERS OF ENDOTHELIAL DYSFUNCTION IN CHRONIC KIDNEY DISEASE PATIENTS. Jhypertension: Abstract
journal of hypertension PP.27.04: ENDOTHELIAL DYSFUNCTION AND ARTERIAL HYPERTENSION IN THE WORKING POPULATION (MERIDIAN STUDY). Jhypertension: Abstract
Curr Vasc Pharmacol. Renalase is Removed by Kidneys and During Dialysis – Excess Related to CKD Complications? PubMed: 25515790
Kidney Blood Press Res Effects of Renin-Angiotensin System Inhibitors on Renal Expression of Renalase in Sprague-Dawley Rats Fed With High Salt Diet PubMed: 26619289
PLoS One Serum Renalase Levels Correlate with Disease Activity in Lupus Nephritis PubMed: 26431044
J Hum Hypertens Plasma renalase concentration before and after radiofrequency renal denervation in patients with resistant hypertension PubMed: 26310184
Nephrology (Carlton) Urinary renalase concentration in patients with preserved kidney function undergoing coronary angiography. pubmed:27778420
Nephrol Dial Transplant PLASMA RENALASE CONCENTRATION BEFORE AND AFTER RADIOFREQUENCY RENAL DENERVATION IN PATIENTS WITH RESISTANT HYPERTENSION: PRELIMINARY RESULTS content:31
Renal Failure Increased serum renalase in hemodialysis patients: is it related to left ventricular hypertrophy? pubmed:27416751
Enhanced biocompatible and hemocompatible nano/micro porous surface as a biological scaffold for functionalizational and biointegrated implants High-performance Liquid Chromatography Measured Metabolites of EndogenousCatecholamines and Their Relations to Chronic Kidney Disease and High Blood Pressure in Heart Transplant Recipients. pubmed:27496485
PLOS ONE Online Haemodiafiltration Improves Inflammatory State in Dialysis Patients: A Longitudinal Study. pubmed:27783636
Advances in Experimental Medicine and Biology Renalase in Children with Glomerular Kidney Diseases chapter:10.1007
Kidney & Blood Pressure Research Plasma Renalase is Not Associated with Blood Pressure and Brachial-Ankle Pulse Wave Velocity in Chinese Adults With Normal Renal Function 452587
BMC nephrology Rs10887800 renalase gene polymorphism influences the level of circulating renalase in patients undergoing hemodialysis but not in healthy controls pubmed:28372594
Nefrología (English Edition) Increased serum renalase in peritoneal dialysis patients: Is it related to cardiovascular disease risk pubmed:28160962
Clinical Interventions in Aging Age influence on renalase and catecholamines concentration in hypertensive patients, including maintained dialysis pubmed:27822026
International Journal of Clinical and Experimental Medicine Aumento de renalasa sérica en pacientes de diálisis peritoneal:¿ está relacionado con el riesgo de enfermedad cardiovascular? 10.1016:j.nefro.2016.11.013
PLoS One Relationship between microRNA-146a expression and plasma renalase levels in hemodialyzed patients pubmed:28614373
Annals of Agricultural and Environmental Medicine Levels of renalase and advanced oxidation protein products with regard to catecholamines in haemodialysed patients pubmed:28954489
Angiology Endocan Concentration in Patients With Primary Hypertension pubmed:29084442
Journal of Hypertension RENALASE PLASMA CONCENTRATION BEFORE AND AFTER RADIOFREQUENCY RENAL DENERVATION IN PATIENTS WITH RESISTANT HYPERTENSION 10.1097/01.hjh.0000523893.07419.ae
International journal of molecular sciences Renalase Protects against Renal Fibrosis by Inhibiting the Activation of the ERK Signaling Pathways pubmed:28448446
Journal of sports medicine and physical fitness Transient changes in serum renalase concentration during long-distance running: The case of an amateur runner under continuous training 10.7600/jpfsm.6.159
Scientific Reports Synergistic effect of renalase and chronic kidney disease on endothelin-1 in patients with coronary artery disease‒a cross-sectional study Pubmed:29743680
Life Sciences Influence of acute exercise on renalase and its regulatory mechanism Pubmed:30056018
Experimental Eye Research Quantitative analysis of hydroxyapatite-binding plasma proteins in genotyped individuals with late-stage age-related macular degeneration Pubmed:29580721
Rapid Communications in Mass Spectrometry A new method for quantitative determination of renalase based on mass‐spectrometry determination of a proteotypic peptide labeled with stable isotopes 10.1002:rcm.8167
International Urology and Nephrology Renalase contributes to protection against renal fibrosis via inhibiting oxidative stress in rats Pubmed:29476431
Pediatric Cardiology Serum Renalase Levels in Adolescents with Primary Hypertension Pubmed:29748702
Postepy w Kardiologii Interwencyjnej Atorvastatin facilitates protection against contrast-induced nephropathy in patients undergoing coronary angiography via humoral mediators rather than … Pubmed:30008773
High urinary kidney injury molecule type 1 level is associated with 12-month adverse cardiovascular events in patients with coronary artery disease publication:324522368
Journal of Clinical Medicine Serum Renalase Levels Are Predicted by Brain-Derived Neurotrophic Factor and Associated with Cardiovascular Events and Mortality after Percutaneous Coronary … Pubmed: 30424498
Archives of disease in childhood GP23 Percutaneous closure of patent ductus arteriosus: single center experience
Archives of disease in childhood GP25 Comparison of associations and outcomes of prenatally and postnatally diagnosed right aortic arch: south wales' experience
Archives of disease in childhood GP24 Serum renalase levels in adolescents with primary hypertension
Disease Markers Association of Plasma Renalase and Left Ventricle Mass Index in Heart Failure Patients Stratified to the Category of the Ejection Fraction: A Pilot Study Pubmed: 31737132
EXPERIMENTAL BIOLOGY AND MEDICINE The role of urinary renalase on early-stage renal damage in Chinese adults with primary hypertension Pubmed: 32102561
FEBS Open Bio Moderate‐intensity exercise increases renalase levels in the blood and skeletal muscle of rats Pubmed: 32053739
current medical research and opinion The partnership between renalase and ejection fraction as a risk factor for increased cardiac remodeling biomarkers in chronic heart failure patients Pubmed: 32297799
Dovepress Association of Serum Renalase Levels and Renalase rs10887800 Polymorphism with Unstable Angina Pectoris Patients Having Metabolic Syndrome Pubmed: 32982355
学位论文 Nikotinamidadenindinukleotid-Blutplasmawerte sind unabhängig vom kalendarischen und vaskulären Alter in einer Normalkohorte aus der Region Halle-Saalekreis
Life Circulating Renalase as Predictor of Renal and Cardiovascular Outcomes in Pre-Dialysis CKD Patients: A 5-Year Prospective Cohort Study 33800219
Edited by: Background: Renalase has been implicated in chronic heart failure (CHF); however, nothing is known about renalase discriminatory ability and prognostic …
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