Multiplex Assay Kit for Angiotensin I Converting Enzyme 2 (ACE2) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

ACEH; ACEII; ACE-II; Peptidyl-Dipeptidase A; ACE-related carboxypeptidase; Angiotensin-converting enzyme homolog; Metalloprotease MPROT15

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

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  • Multiplex Assay Kit for Angiotensin I Converting Enzyme 2 (ACE2) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Angiotensin I Converting Enzyme 2 (ACE2) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Angiotensin I Converting Enzyme 2 (ACE2) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMB886Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Angiotensin I Converting Enzyme 2 (ACE2) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiotensin I Converting Enzyme 2 (ACE2) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-94 91
EDTA plasma(n=5) 89-97 92
heparin plasma(n=5) 95-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiotensin I Converting Enzyme 2 (ACE2) ,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 Angiotensin I Converting Enzyme 2 (ACE2) ,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 Angiotensin I Converting Enzyme 2 (ACE2) ,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) 80-99% 98-105% 92-105% 80-103%
EDTA plasma(n=5) 91-101% 94-103% 80-98% 95-102%
heparin plasma(n=5) 81-90% 81-95% 82-89% 84-97%

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#:ACE2) 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 Angiotensin I Converting Enzyme 2 (ACE2) ,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

  • Angiotensin-Converting Enzyme 2 Overexpression Remarkably Ameliorated Glomerular Injury in a Rat Model of Diabetic Nephropathy: A Comparison with ACE InhibitionMolMed: 10_11_liu
  • Plasma Levels of Angiotensin-Converting Enzymes 1 and 2 and AGTR2 (T1247G and A5235G) Gene Polymorphisms Are Associated to Breast Cancer Progression.Scirp: Source
  • ACE2 activation by xanthenone prevents leptin-induced increases in blood pressure and proteinuria during pregnancy in Sprague-Dawley ratsPubmed:25205467
  • Angiotensin converting enzymes in patients with acute respiratory distress syndromeContent: 3
  • Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosisS0021967317311858
  • Relationship between genetic variants of ACE2 gene and circulating levels of ACE2 and its metabolites.pubmed:28895159
  • Lipoxin A attenuates LPS-induced acute lung injury via activation of the ACE2-Ang-(1-7)-Mas axisPubmed:29969931
  • Impact of angiotensin-converting enzyme 2 levels on postoperative pneumonia after esophagectomyPubmed:29506841
  • Could cardioprotective effect of ACE2 activator “diminazene aceturate” is more potent than ACE inhibitor “Enalapril” on acute myocardial infarction in rats?Pubmed: 30840489
  • Angiotensin-converting enzymes in acute respiratory distress syndrome
  • AVE 0991 Attenuates Pyroptosis and Liver Damage after Heatstroke by Inhibiting the ROS-NLRP3 Inflammatory Signalling Pathway
  • Relationship between circulating levels of angiotensin-converting enzyme 2-angiotensin-(1–7)-MAS axis and coronary heart diseasePubmed: 31359146
  • Alteration and association between serum ACE2/angiotensin (1-7)/Mas axis and oxidative stress in chronic kidney disease: A pilot studyPubmed: 32756181
  • Equilibrium Angiotensin Metabolite Profiling in Patients with Acute Respiratory Distress Syndrome Indicates Angiotensin Converting Enzyme InhibitionPubmed: 32628511
  • Determinants of soluble angiotensin-converting enzyme 2 concentrations in adult patients with complex congenital heart disease33280062
  • Pattern of circulating SARS©\CoV©\2©\specific antibody©\secreting and memory B©\cell generation in patients with acute COVID©\1933552508
  • Novel transgenic mice with Cre-dependent co-expression of GFP and human ACE2: a safe tool for study of COVID-19 pathogenesis33855640
  • Different Neutralization Sensitivity of SARS-CoV-2 Cell-to-Cell and Cell-Free Modes of Infection to Convalescent Sera
  • Immunoreactivity of the SARS-CoV-2 entry proteins ACE-2 and TMPRSS-2 in murine models of hormonal manipulation, ageing, and cardiac injury34907257
  • Angiotensin‐converting enzyme 2 and Transmembrane protease serine 2 in female and male patients with end‐stage kidney diseasePubmed:35366343
  • Ramipril Reduces Acylcarnitines and Distinctly Increases Angiotensin-Converting Enzyme 2 Expression in Lungs of RatsPubmed:35448480

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