ELISA Kit for Matrix Gla Protein (MGP)

MGLAP; GIG36; Cell growth-inhibiting gene 36 protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Matrix Gla Protein (MGP).
No significant cross-reactivity or interference between Matrix Gla Protein (MGP) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Matrix Gla Protein (MGP) and the recovery rates were calculated by comparing the measured value to the expected amount of Matrix Gla Protein (MGP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-97 82
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 87-98 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Matrix Gla Protein (MGP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Matrix Gla Protein (MGP) 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 Matrix Gla Protein (MGP) 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) 93-101% 96-105% 96-105% 94-101%
EDTA plasma(n=5) 85-101% 79-92% 78-105% 93-103%
heparin plasma(n=5) 83-102% 94-104% 88-95% 87-95%

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
Nephrology Dialysis Transplantation Magnesium prevents phosphate-induced calcification in human aortic vascular smooth muscle cells PubMed: PMC3611891
Maturitas. Bone metabolism regulators and arterial stiffness in postmenopausal women Pubmed: 23916080
The Egyptian Rheumatologist Osteoprotegerin (OPG) and Matrix Gla protein (MGP) in rheumatoid arthritis patients: Relation to disease activity ScienceDirect: S1110116414000076
Toxicology Reports Canola and hydrogenated soybean oils accelerate ectopic bone formation induced by implantation of bone morphogenetic protein in mice Researchgate:Source
Lipids in Health and Disease Markers of increased cardiovascular risk in patients with chronic kidney disease Biomedcentral:Source
Human &amp; Veterinary Medicine The behavior of circulating matrix Gla protein, matrix metalloproteinase-9 and nitrotyrosine in patients with varicose veins Hvm:Source
Pol Arch Med Wewn Effect of Vitamin K2 on progression of atherosclerosis and vascular calcification in nondialyzed patients with chronic kidney disease stages 3-5 PubMed: 26176325
Lipids Health Dis. Markers of increased atherosclerotic risk in patients with chronic kidney disease: a preliminary study pmc:PMC4739105
Kidney Blood Press Res. Plasma Desphospho-Uncarboxylated Matrix Gla Protein as a Marker of Kidney Damage and Cardiovascular Risk in Advanced Stage of Chronic Kidney Disease Pubmed:27100101
Archives of Oral Biology Matrix-Gla Protein rs4236 [A/G] gene polymorphism and serum and GCF levels of MGP in patients with subgingival dental calculus pubmed:27348051
Cell Death & Disease Mesenchymal stromal cells-derived matrix Gla protein contribute to the alleviation of experimental colitis Pubmed:29880866
Urology High concentration of calcium promotes mineralization in NRK52E cells via inhibiting the expression of matrix Gla protein Pubmed:29935264
Clinica Chimica Acta Serum total matrix Gla protein: Reference interval in healthy adults and variations in patients with vascular and osteoarticular diseases Pubmed: 30597137
Biological Trace Element Research Altered Mineral Metabolism and Disequilibrium Between Calcification Promoters and Inhibitors in Chronic Hemodialysis Patients Pubmed: 30847765
Renal Failure Relationship of matrix Gla protein and vitamin K with vascular calcification in hemodialysis patients Pubmed: 31538831
Heliyon Matrix Gla protein maintains normal and malignant hematopoietic progenitor cells by interacting with bone morphogenetic protein-4 Pubmed: 32322728
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