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

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)

Specificity

This assay has high sensitivity and excellent specificity for detection of Fibulin 3 (FBLN3) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Fibulin 3 (FBLN3) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

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) 78-92 86
EDTA plasma(n=5) 78-89 82
heparin plasma(n=5) 81-90 86

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) 87-96% 85-99% 79-101% 78-101%
EDTA plasma(n=5) 86-96% 78-93% 96-104% 82-102%
heparin plasma(n=5) 79-104% 93-101% 86-101% 96-104%

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.

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Magazine Citations
1 Fibulin-3 as a Blood and Effusion Biomarker for Pleural Mesothelioma PubMed: PMC3761217
Anticancer Res. YKL-40 and mesothelin in the blood of patients with malignant mesothelioma, lung cancer and asbestosis. Pubmed: 24324091
Oncology Reports Role of fibulin-3 in lung cancer: In vivo and in vitro analyses Spandidos-publications: Source
Current Pulmonology Reports Discovery of new biomarkers for malignant mesothelioma Springer:Source
United States Patent Application 20140378336 DIAGNOSIS OF CELL PROLIFERATIVE DISEASES Freepatentsonline:Source
Thorax. Comparison of fibulin-3 and mesothelin as markers in malignant mesothelioma Pubmed:Pmc4174124
Egyptian Journal of Chest Diseases and Tuberculosis Value of plasma and pleural effusion fibulin-3 levels in the diagnosis of malignant pleural mesothelioma effusions Pubmed:23050525
Radiol Oncol Fibulin-3 as a biomarker of response to treatment in malignant mesothelioma PubMed: 26401134
British Journal of Cancer Fibulin-3 levels in malignant pleural mesothelioma are associated with prognosis but not diagnosis PubMed: 26263483
J Thorac Oncol. Plasma Biomarker Enrichment of Clinical Prognostic Indices in Malignant Pleural Mesothelioma. Pubmed:26903362
BMJ Open Diagnostic and Prognostic Biomarkers in the Rational Assessment of Mesothelioma (DIAPHRAGM) study: protocol of a prospective, multicentre, observational study pubmed:27884852
25 HMGB1 and its hyperacetylated isoform are sensitive and specific serum biomarkers to detect asbestos exposure and to identify mesothelioma patients pubmed:26733616
BMJ Journals Diagnostic and Prognostic Biomarkers in the Rational Assessment of Mesothelioma (DIAPHRAGM) study: protocol of a prospective, multicentre, observational … e013324.abstract
Anticancer Res. Comparison of the Diagnostic Performance of Fibulin-3 and Mesothelin in Patients with Pleural Effusions from Malignant Mesothelioma. pubmed:28314308
Dis Markers. A Novel Panel of Serum Biomarkers for MPM Diagnosis. pubmed:28348450
The Knee Association of fibulin-3 concentrations with the presence and severity of knee osteoarthritis: A cross-sectional study pubmed:29195846
Disease markers Plasma Fibulin-3 as a Potential Biomarker for Patients with Asbestos-Related Diseases in the Han Population pubmed:29200597
Molecular Medicine Reports Placental protein 14 as a potential biomarker for diagnosis of preterm premature rupture of membranes Pubmed:29749501
Oncotarget The matrix protein Fibulin-3 promotes KISS1R induced triple negative breast cancer cell invasion Pubmed:30046386
Advances in Clinical and Experimental Medicine Fibulin-3 and other cartilage metabolism biomarkers in relationship to calprotectin (MRP8/14) and disease activity in rheumatoid arthritis patients treated with … 68362.pdf
PLoS One Prediction of pneumoconiosis by serum and urinary biomarkers in workers exposed to asbestos-contaminated minerals Pubmed: 30946771
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
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