Multiplex Assay Kit for Neprilysin (CD10) ,etc. by FLIA (Flow Luminescence Immunoassay)

NEP; MME; CALLA; SFE; Atriopeptidase; Enkephalinase; Skin fibroblast elastase; Common Acute Lymphoblastic Leukemia Antigen; Membrane Metallo-Endopeptidase

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Neprilysin (CD10) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Neprilysin (CD10) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Neprilysin (CD10) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Neprilysin (CD10) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 79-89 83
EDTA plasma(n=5) 88-95 92
heparin plasma(n=5) 99-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Neprilysin (CD10) ,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 Neprilysin (CD10) ,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 Neprilysin (CD10) ,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) 80-102% 78-90% 92-105% 90-104%
EDTA plasma(n=5) 83-91% 84-92% 91-98% 97-105%
heparin plasma(n=5) 92-99% 79-93% 78-95% 89-96%

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#:CD10) 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
13 Elevated Plasma levels of Neutral Endpeptidase in Hypertension Patients Treated with Angiotensin Converting Enzyme Inhibitors Ahajournals:Source
Biol Pharm Bull Effect of High Glucose Levels on Amyloid β Production in Retinas of Spontaneous Diabetes Mellitus Otsuka Long-Evans Tokushima Fatty Rats PubMed: 25832640
Eur J Heart Fail.  Soluble neprilysin does not correlate with outcome in heart failure with preserved ejection fraction Pubmed:26725876
8 A Test in Context: Neprilysin: Function, Inhibition, and Biomarker. pubmed:27491909
European Journal of Heart Failure Effects of sacubitril/valsartan on neprilysin targets and the metabolism of natriuretic peptides in chronic heart failure: a mechanistic clinical study Pubmed: 30520545
European Journal of Heart Failure Increased granulocyte membrane neprilysin (CD10) expression is associated with better prognosis in heart failure Pubmed: 30828920
Clinical Research in Cardiology Soluble neprilysin, NT-proBNP, and growth differentiation factor-15 as biomarkers for heart failure in dialysis patients (SONGBIRD) Pubmed: 32002632
Neurotoxicology Involvement of hippocampal agmatine in β1-42 amyloid induced memory impairment, neuroinflammation and BDNF signaling disruption in mice Pubmed: 32522471
European Heart Journal P1620 Association of granulocyte neprilysin (CD10) expression with prognosis in heart failure with reduced ejection fraction
Clin Biochem Finding a reliable assay for soluble neprilysin Pubmed:35331754
Scientific Reports The potential role of serum tau protein (MAPT), neuronal cell adhesion molecule (NrCAM) and neprilysin (NEP) in neurodegenerative disorders development in …
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MAB785Hu24 Monoclonal Antibody to Neprilysin (CD10) WB; IHC; ICC; IP.
MAB785Hu22 Monoclonal Antibody to Neprilysin (CD10) WB; IHC; ICC; IP.
MAB785Hu25 Monoclonal Antibody to Neprilysin (CD10) WB; IHC; ICC; IP.
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