Multiplex Assay Kit for Endothelial Cell Specific Molecule 1 (ESM1) ,etc. by FLIA (Flow Luminescence Immunoassay)

Endocan

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Endothelial Cell Specific Molecule 1 (ESM1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Endothelial Cell Specific Molecule 1 (ESM1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Endothelial Cell Specific Molecule 1 (ESM1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Endothelial Cell Specific Molecule 1 (ESM1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 102
EDTA plasma(n=5) 79-101 94
heparin plasma(n=5) 79-99 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Endothelial Cell Specific Molecule 1 (ESM1) ,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 Endothelial Cell Specific Molecule 1 (ESM1) ,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 Endothelial Cell Specific Molecule 1 (ESM1) ,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) 89-97% 91-105% 89-98% 79-95%
EDTA plasma(n=5) 98-105% 79-98% 88-96% 84-98%
heparin plasma(n=5) 87-95% 79-96% 93-103% 89-97%

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#:ESM1) 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
Urology Detection on dynamic changes of endothelial cell specific molecule-1 in acute rejection after renal transplantation PubMed: 22608803
Microvasc Res. Endocan is useful biomarker of survival and severity in sepsis. Pubmed: 24769132
J Matern Fetal Neonatal Med. Endocan, a putative endothelial cell marker, is elevated in preeclampsia, decreased in acute pyelonephritis, and unchanged in other obstetrical syndromes Pubmed:25211383
Microvasc Res. Endocan is useful biomarker of survival and severity in sepsis Pubmed:24769132
Angiology Endocan Levels and Subclinical Atherosclerosis in Patients With Systemic Lupus Erythematosus PubMed: 26614790
Archives of Medical Science - Atherosclerotic Diseases Assessment of subclinical cardiac damage in chronic plaque psoriasis patients: a case control study Journal:-102
Int Immunopharmacol.  Inhibition of endocan attenuates monocrotaline-induced connective tissue disease related pulmonary arterial hypertension. pubmed:27912147
Oncotarget Diagnostic and prognostic values of endothelial-cell-specific molecule-1 with malignant pleural effusions in patients with non-small cell lung cancer pubmed:28514746
Biomedical and Pharmacology Journal Changes in Endocan Levels and Blood Coagulation in HIV Infection DOI: 10.13005/bpj/1082
Life Sciences A novel role of endocan in alleviating LPS-induced acute lung injury Pubmed:29627442
International Urology and Nephrology Is the plasma endocan level a reliable predictor for the severity of erectile dysfunction? Pubmed:30054863
The Open Rheumatology Journal Serum Level of Endothelial Cell-Specific Molecule-1 (ESM-1) as a New Potential Biomarker for Rheumatoid Arthritis Disease Activity Doi: 10.2174/1874312901812010189
Molecular Medicine Reports Downregulation of ENDOCAN in myeloid leukemia cells inhibits proliferation and promotes apoptosis by suppressing nuclear factor‑κB activity Pubmed: 30816462
Biomarkers in Medicine Endothelial biomarkers in the light of new sepsis definition Pubmed: 30920847
Experimental and Therapeutic Medicine Risk stratification and prognostic evaluation of endothelial cell‑specific molecule1, von Willebrand factor, and a disintegrin‑like and metalloprotease with …
Turkish Journal of Medical Sciences A novel marker relationship between carotid intima–media thickness and disease activity score-28 in patients with rheumatoid arthritis: human endothelial cell … Pubmed: 31655533
Biomarkers Role of Ascitic Endocan Levels in the Diagnosis of Spontaneous Bacterial Peritonitis in Decompensated Cirrhosis Pubmed: 32364003
J Obstet Gynaecol Res Comparison of the maternal serum endocan levels in preterm premature rupture of membrane and normal pregnancy 34109715
turkish journal clinics laboratory The relationship between serum endocan levels with the presence of contrast-induced nephropathy in patients undergoing coronary angiography
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