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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)
- Product No.LMB785Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cerebrospinal fluid, cell culture supernates and other biological fluids
- Test MethodDouble-antibody Sandwich
- Assay Length3.5h
- Detection Range1.95-2000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.65 pg/mL.
- DownloadInstruction Manual
- UOM 8Plex 7Plex 6Plex 5Plex 4Plex 3Plex 2Plex1Plex
- FOB
US$ 415
US$ 431
US$ 455
US$ 487
US$ 519
US$ 567
US$ 638
Result
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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 … |