ELISA Kit for Matrix Gla Protein (MGP) Homo sapiens (Human) Sandwich ELISA

MGLAP; GIG36; Cell growth-inhibiting gene 36 protein

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  • ELISA Kit for Matrix Gla Protein (MGP) Packages (Simulation)
  • ELISA Kit for Matrix Gla Protein (MGP) Packages (Simulation)
  • ELISA Kit for Matrix Gla Protein (MGP) Results demonstration
  • SEB477Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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.

ELISA Kit for Matrix Gla Protein (MGP)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Matrix Gla Protein (MGP). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Matrix Gla Protein (MGP). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Matrix Gla Protein (MGP), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Matrix Gla Protein (MGP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Magnesium prevents phosphate-induced calcification in human aortic vascular smooth muscle cellsPubMed: PMC3611891
  • Bone metabolism regulators and arterial stiffness in postmenopausal womenPubmed: 23916080
  • Osteoprotegerin (OPG) and Matrix Gla protein (MGP) in rheumatoid arthritis patients: Relation to disease activityScienceDirect: S1110116414000076
  • Canola and hydrogenated soybean oils accelerate ectopic bone formation induced by implantation of bone morphogenetic protein in miceResearchgate:Source
  • Markers of increased cardiovascular risk in patients with chronic kidney diseaseBiomedcentral:Source
  • The behavior of circulating matrix Gla protein, matrix metalloproteinase-9 and nitrotyrosine in patients with varicose veinsHvm:Source
  • Effect of Vitamin K2 on progression of atherosclerosis and vascular calcification in nondialyzed patients with chronic kidney disease stages 3-5PubMed: 26176325
  • Markers of increased atherosclerotic risk in patients with chronic kidney disease: a preliminary studypmc:PMC4739105
  • Plasma Desphospho-Uncarboxylated Matrix Gla Protein as a Marker of Kidney Damage and Cardiovascular Risk in Advanced Stage of Chronic Kidney DiseasePubmed:27100101
  • Matrix-Gla Protein rs4236 [A/G] gene polymorphism and serum and GCF levels of MGP in patients with subgingival dental calculuspubmed:27348051
  • Mesenchymal stromal cells-derived matrix Gla protein contribute to the alleviation of experimental colitisPubmed:29880866
  • High concentration of calcium promotes mineralization in NRK52E cells via inhibiting the expression of matrix Gla proteinPubmed:29935264
  • Serum total matrix Gla protein: Reference interval in healthy adults and variations in patients with vascular and osteoarticular diseasesPubmed: 30597137
  • Altered Mineral Metabolism and Disequilibrium Between Calcification Promoters and Inhibitors in Chronic Hemodialysis PatientsPubmed: 30847765
  • Relationship of matrix Gla protein and vitamin K with vascular calcification in hemodialysis patientsPubmed: 31538831
  • Matrix Gla protein maintains normal and malignant hematopoietic progenitor cells by interacting with bone morphogenetic protein-4Pubmed: 32322728

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