Multiplex Assay Kit for Neutrophil Elastase (NE) ,etc. by FLIA (Flow Luminescence Immunoassay) Canis familiaris; Canine (Dog) Multiplex ELISA

ELANE; HLE; HNE; ELA2; PMN-E; Neutrophil Elastase; Medullasin; Polymorphonuclear Leukocyte Elastase; Bone Marrow Serine Protease; Leukocyte Elastase; PMN Elastase

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

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  • Multiplex Assay Kit for Neutrophil Elastase (NE) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Neutrophil Elastase (NE) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Neutrophil Elastase (NE) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA181Ca.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-93 85
EDTA plasma(n=5) 90-97 93
heparin plasma(n=5) 78-88 82

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Neutrophil Elastase (NE) ,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 Elastase (NE) ,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 Elastase (NE) ,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) 83-98% 78-97% 87-94% 79-89%
EDTA plasma(n=5) 81-89% 89-97% 80-88% 78-104%
heparin plasma(n=5) 81-88% 79-93% 80-98% 82-96%

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#:NE) 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.

Multiplex Assay Kit for Neutrophil Elastase (NE) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Long-Term Leukocyte Filtration Should Be Avoided during Extracorporeal CirculationHindawi: 612848
  • A dietary supplement improves facial photoaging and skin sebum, hydration and tonicity modulating serum fibronectin, neutrophil elastase 2, hyaluronic acid and carbonylated proteinsPubmed:25732262
  • The neutrophil elastase inhibitor, sivelestat, attenuates sepsis-related kidney injury in ratspubmed:27430552
  • Xuebijing injection treatment inhibits vasopermeability and reduces fluid requirements in acanine burn model.pubmed:28070608
  • Inhibition of ALDH2 protects PC12 cells against formaldehyde-induced cytotoxicity: involving the protection of hydrogen sulphide.pubmed:28251688
  • Targeted regulation of sympathetic activity in paraventricular nucleus reduces inducible ventricular arrhythmias in rats after myocardial infarction10.1016:j.jjcc.2018.06.003
  • Angiotensin II Type 1a Receptors in the Subfornical Organ Modulate Neuroinflammation in the Hypothalamic Paraventricular Nucleus in Heart Failure RatsPubmed:29684507
  • Repetitive transcranial magnetic stimulation inhibits Sirt1/MAO-A signaling in the prefrontal cortex in a rat model of depression and cortex-derived astrocytesPubmed:28948423
  • The depressive-like behaviors of chronic unpredictable mild stress-treated mice, ameliorated by Tibetan medicine Zuotai: involvement in the hypothalamic …Pubmed:29379286
  • Possible mechanisms of prenatal cold stress induced-anxiety-like behavior depression in offspring ratsPubmed: 30423388
  • Role of melatonin in sleep deprivation‐induced intestinal barrier dysfunction in micePubmed: 30929267
  • Effect of antimicrobial peptides HNP‐1 and hBD‐1 on Staphylococcus aureus strains in vitro and in vivoPubmed: 31313350
  • Erythromycin suppresses neutrophil extracellular traps in smoking-related chronic pulmonary inflammationPubmed: 31515489
  • Co-Administration of Curcumin and Artepillin C Induces Development of Brown-Like Adipocytes in Association with Local Norepinephrine Production by Alternatively …Pubmed: 31474682
  • MiR-608 Exerts Anti-inflammatory Effects by Targeting ELANE in MonocytesPubmed: 31749032
  • Neutrophil Extracellular Traps Associate with Clinical Stages in Breast CancerPubmed: 31656990
  • Inhibition of microRNA-146a attenuated heart failure in myocardial infarction ratsPubmed: 31763669
  • Neutrophil Elastase Inhibitors Suppress Oxidative Stress in Lung during Liver TransplantationPubmed: 31871555
  • Levels of pro-and anti-inflammatory cytokines in cystic fibrosis patients with or without gingivitisPubmed: 31927460
  • Neurotransmitter system aberrations in patients with drug addictionPubmed: 32804296
  • Corneal epithelial injury-induced norepinephrine promotes Pseudomonas aeruginosa keratitisPubmed: 32376471
  • Hyperinsulinemia rather than insulin resistance itself induces blood pressure elevation in high fat diet-fed ratsPubmed: 32349626
  • Macrophage Extracellular Traps Aggravate Iron Overload©\Related Liver Ischemia/Reperfusion Injury33959955
  • Zisheng Shenqi Decoction Ameliorates Monosodium Urate-Mediated Gouty Arthritis in Rats via Promotion of Autophagy through the AMPK/mTOR Signaling?¡­33505502
  • Depression of Non-Neuronal Cholinergic System May Play a Role in Co-Occurrence of Subjective Daytime Sleepiness and Hypertension in Patients with …34934375
  • Transcutaneous Electrical Acustimulation Improved the Quality of Life in Patients With Diarrhea-Irritable Bowel SyndromePubmed:35088760
  • A Novel Role of Tinospora Cordifolia in Amelioration of Cancer-Induced Systemic Deterioration By Taming Neutrophil Infiltration and Hyperactivation

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