Multiplex Assay Kit for Fibulin 3 (FBLN3) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

EFEMP1; DHRD; DRAD; FBNL; MLVT; MTLV; S1-5; EGF-Containing Fibulin-Like Extracellular Matrix Protein 1; Fibrillin-Like; Extracellular protein S1-5

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

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  • Multiplex Assay Kit for Fibulin 3 (FBLN3) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Fibulin 3 (FBLN3) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Fibulin 3 (FBLN3) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMF422Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Fibulin 3 (FBLN3) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Fibulin 3 (FBLN3) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-105 101
EDTA plasma(n=5) 83-97 86
heparin plasma(n=5) 83-91 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibulin 3 (FBLN3) ,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 Fibulin 3 (FBLN3) ,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 Fibulin 3 (FBLN3) ,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) 97-104% 81-97% 78-90% 85-99%
EDTA plasma(n=5) 84-98% 81-98% 94-105% 97-105%
heparin plasma(n=5) 83-96% 78-101% 86-103% 97-105%

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#:FBLN3) 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 Fibulin 3 (FBLN3) ,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

  • Fibulin-3 as a Blood and Effusion Biomarker for Pleural MesotheliomaPubMed: PMC3761217
  • YKL-40 and mesothelin in the blood of patients with malignant mesothelioma, lung cancer and asbestosis.Pubmed: 24324091
  • Role of fibulin-3 in lung cancer: In vivo and in vitro analysesSpandidos-publications: Source
  • Discovery of new biomarkers for malignant mesotheliomaSpringer:Source
  • DIAGNOSIS OF CELL PROLIFERATIVE DISEASESFreepatentsonline:Source
  • Comparison of fibulin-3 and mesothelin as markers in malignant mesotheliomaPubmed:Pmc4174124
  • Value of plasma and pleural effusion fibulin-3 levels in the diagnosis of malignant pleural mesothelioma effusionsPubmed:23050525
  • Fibulin-3 as a biomarker of response to treatment in malignant mesotheliomaPubMed: 26401134
  • Fibulin-3 levels in malignant pleural mesothelioma are associated with prognosis but not diagnosisPubMed: 26263483
  • Plasma Biomarker Enrichment of Clinical Prognostic Indices in Malignant Pleural Mesothelioma.Pubmed:26903362
  • Diagnostic and Prognostic Biomarkers in the Rational Assessment of Mesothelioma (DIAPHRAGM) study: protocol of a prospective, multicentre, observational study pubmed:27884852
  • HMGB1 and its hyperacetylated isoform are sensitive and specific serum biomarkers to detect asbestos exposure and to identify mesothelioma patientspubmed:26733616
  • Diagnostic and Prognostic Biomarkers in the Rational Assessment of Mesothelioma (DIAPHRAGM) study: protocol of a prospective, multicentre, observational …e013324.abstract
  • Comparison of the Diagnostic Performance of Fibulin-3 and Mesothelin in Patients with Pleural Effusions from Malignant Mesothelioma.pubmed:28314308
  • A Novel Panel of Serum Biomarkers for MPM Diagnosis.pubmed:28348450
  • Association of fibulin-3 concentrations with the presence and severity of knee osteoarthritis: A cross-sectional studypubmed:29195846
  • Plasma Fibulin-3 as a Potential Biomarker for Patients with Asbestos-Related Diseases in the Han Populationpubmed:29200597
  • Placental protein 14 as a potential biomarker for diagnosis of preterm premature rupture of membranesPubmed:29749501
  • The matrix protein Fibulin-3 promotes KISS1R induced triple negative breast cancer cell invasionPubmed:30046386
  • Fibulin-3 and other cartilage metabolism biomarkers in relationship to calprotectin (MRP8/14) and disease activity in rheumatoid arthritis patients treated with …68362.pdf
  • Prediction of pneumoconiosis by serum and urinary biomarkers in workers exposed to asbestos-contaminated mineralsPubmed: 30946771

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