Multiplex Assay Kit for Osteonectin (ON) ,etc. by FLIA (Flow Luminescence Immunoassay)

SPARC; BM-40; Secreted Protein,Acidic,Cysteine-Rich; Basement-membrane protein 40; Secreted protein acidic and rich in cysteine

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-101 90
EDTA plasma(n=5) 91-99 96
heparin plasma(n=5) 96-104 101
sodium citrate plasma(n=5) 95-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Osteonectin (ON) ,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 Osteonectin (ON) ,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 Osteonectin (ON) ,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) 90-98% 84-103% 79-93% 98-105%
EDTA plasma(n=5) 83-98% 86-101% 78-99% 93-101%
heparin plasma(n=5) 84-92% 90-103% 79-101% 96-105%
sodium citrate plasma(n=5) 98-105% 90-101% 87-105% 92-105%

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#:ON) 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
J Mol Med (Berl) Growth hormone replacement therapy regulates microRNA-29a and targets involved in insulin resistance PubMed: 26199111
Muscle Nerve Effect of resistance ladder training on SPARC expression in skeletal muscle of hindlimb immobilized rats PubMed: 26467437
Medicine (Baltimore) An Attempt to Evaluate Selected Aspects of “Bone–Fat Axis” Function in Healthy Individuals and Patients With Pancreatic Cancer PubMed: 26266370
British Journal of Cancer A proteomics-based approach identifies secreted protein acidic and rich in cysteine as a prognostic biomarker in malignant pleural mesothelioma Pubmed:26889976
Journal of diabetes and its complications Associations between FGF21, osteonectin and bone turnover markers in type 2 diabetic patients with albuminuria. pubmed:27916484
American journal of the medical sciences Association of Bone Metabolic Markers With Diabetic Retinopathy and Diabetic Macular Edema in Elderly Chinese Individuals With Type 2 Diabetes Mellitus pubmed:29078839
Translational Research in Oral Oncology Testican 1 (SPOCK1) and protein tyrosine phosphatase, receptor type S (PTPRS) show significant increase in saliva of tobacco users with oral cancer Doi: 10.1177/2057178X18800534
[ENGINEERING OF BIOMATERIALS Osteoconductive potential of PLGA/bioglass composite biomaterials
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