Multiplex Assay Kit for Copeptin (CPP) ,etc. by FLIA (Flow Luminescence Immunoassay)

C-Terminal Provasopressin; Vasopressin-Neurophysin 2-Copeptin

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-104 101
EDTA plasma(n=5) 78-92 88
heparin plasma(n=5) 95-104 98
sodium citrate plasma(n=5) 90-104 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Copeptin (CPP) ,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 Copeptin (CPP) ,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 Copeptin (CPP) ,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) 84-96% 87-95% 92-104% 85-95%
EDTA plasma(n=5) 96-104% 98-105% 85-102% 95-103%
heparin plasma(n=5) 99-105% 98-105% 85-99% 80-94%
sodium citrate plasma(n=5) 78-98% 89-99% 93-101% 98-105%

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#:CPP) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Med-Science. Copeptin: May it be a novel biomarker for insulin theraphy in diabetes? An animal study. Ejmanager: Source
Psychiatry and Clinical Neurosciences Serum copeptin in children exposed to maltreatment Pubmed:27278269
United States Patent Application Methods for Diagnosis and Treatment of Concussion or Brain Injury 2016:0103140
J Physiol Role of enteroendocrine L‐cells in arginine vasopressin‐mediated inhibition of colonic anion secretion Pubmed:27037495
International Journal of Clinical and Experimental Medicine Can hypoxia enhance sexual arousal?-Molecular-biological analysis of the hypothalamus in male rats placed with oestrous female rats under hypoxic … files:ijcem0031358.pdf
Front Physiol Orthostatic Challenge Shifts the Hemostatic System of Patients Recovered from Stroke toward Hypercoagulability. pubmed:28223937
Kidney Blood Pressure Research N-Terminal Prohormone of Brain Natriuretic Peptide but not C-Terminal Pre-Pro Vasopressin (Copeptin) Level is Associated with the Response to Antihypertensive … pubmed:29190613
Hindawi Publishing Corporation Early Prediction of Preeclampsia journals:297397
Journal of Urology Evaluation of Urinary Aquaporin 2 and Plasma Copeptin as Biomarkers of Effectiveness of Desmopressin Acetate for the Treatment of Monosymptomatic Nocturnal Enuresis. pubmed:28457803
American Journal of Physiology-Renal Physiology Experimental Heat Stress Nephropathy and Liver Injury are Improved by Allopurinol Pubmed:29667911
American Journal of Physiology-Regulatory Integrative and Comparative Physiology Angiotensin AT Receptors Expressed in Vasopressin-Producing Cells of the Supraoptic Nucleus Contribute to the Osmotic Control of Vasopressin Pubmed:29364700
BMC Nephrology Rehydration with fructose worsens dehydration-induced renal damage Pubmed:30005632
Experimental and Therapeutic Medicine Copeptin level in the early prediction of cardiorenal syndrome in rats Pubmed:30112047
中国病理生理杂志 和肽素水平对大鼠心肾综合征的早期预测价值 filename:ZBLS201802013
Hypertension Abstract P311: Static Urine Osmolality With Elevated First Trimester Urine Copeptin in Human Preeclampsia
Journal of Pediatric Urology Assessment of serum levels of copeptin and corticotropin-releasing factor in children with monosymptomatic and nonmonosymptomatic nocturnal enuresis
experimental physiology Cognitive performance is associated with cerebral oxygenation and peripheral oxygen saturation, but not plasma catecholamines, during graded normobaric hypoxia
international urology and nephrology Dialysate copeptin and peritoneal transport in incident peritoneal dialysis patients Pubmed: 31187425
American Journal of Physiology-renal Physiology The contribution of collecting duct NOS1 to the concentrating mechanisms in male and female mice Pubmed: 31241990
american journal of physiology-renal physiology The increase of core temperature affected the progression of kidney injury by repeated heat stress exposure Pubmed: 31390229
Scientific Reports Deletion of the serine protease CAP2/Tmprss4 leads to dysregulated renal water handling upon dietary potassium depletion Pubmed: 31863073
BMC Nephrology Copeptin is independently associated with vascular calcification in chronic kidney disease stage 5 Pubmed: 32033584
Archives of Biochemistry and Biophysics Micro RNA 146a gene variant/TNF-α/IL-6/IL-1 β; A cross-link axis inbetween oxidative stress, endothelial dysfunction and neuro-inflammation in acute ischemic stroke … Pubmed: 31738890
Ann Nutr Metab Copeptin and Nesfatin-1 Are Interrelated Biomarkers with Roles in the Pathogenesis of Insulin Resistance in Chinese Children with Obesity Pubmed: 33027789
Biomolecules . Multimarker Approach to Identify Patients with Coronary Artery Disease at High Risk for Subsequent Cardiac Adverse Events: The Multi-Biomarker Study Pubmed: 32549327
Arch Med Res Relation of Copeptin with Diabetic and Renal Function Markers Among Patients with Diabetes Mellitus Progressing Towards Diabetic Nephropathy Pubmed: 32505416
河北医科大学学报 重组人脑利钠肽治疗慢性肺心病急性加重期的临床效果及对5-HT 浓度的影响
Scientific reports Validity of different copeptin assays in the differential diagnosis of the polyuria-polydipsia syndrome 33980941
International Journal of Women¡¯s Health and Reproduction Sciences Are the Circulating Levels of Copeptin and Fibronectin Dysregulated in Preeclamptic South African Black Women?
Dicle T?p Dergisi Copeptin levels are more elevated and associated with cardiovascular events in the unity of chronic ischemic heart and chronic kidney diseases
High serum copeptin may be a marker of an increased carotid intima-media thickness in asymptomatic patients with type 1 diabetes.
Nutrients The Speed of Ingestion of a Sugary Beverage Has an Effect on the Acute Metabolic Response to Fructose
Acta Physiol (Oxf) Dietary K+ acts as a genuine diuretic Pubmed:34984847
Anal Chim Acta Spectrally multiplexed assay using gap enhanced nanoparticle for detection of a myocardial infarction biomarker panel Pubmed:35190126
J Cell Physiol Arcuate NPY is involved in salt‐induced hypertension via modulation of paraventricular vasopressin and brain‐derived neurotrophic factor Pubmed:35312067
Children Copeptin Concentrations in Plasma of Healthy Neonates in Relation to Water–Electrolyte Homeostasis in the Early Adaptation Period Pubmed:35327814
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