ELISA Kit for Fibulin 3 (FBLN3)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 95-102 99
EDTA plasma(n=5) 89-101 93
heparin plasma(n=5) 96-104 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibulin 3 (FBLN3) 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) 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) 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) 99-105% 86-104% 78-94% 91-99%
EDTA plasma(n=5) 91-98% 78-94% 87-94% 90-98%
heparin plasma(n=5) 92-99% 98-105% 89-96% 96-103%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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
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