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基質(zhì)Gla蛋白(MGP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Matrix Gla Protein (MGP)

MGLAP; GIG36; Cell growth-inhibiting gene 36 protein

  • 基質(zhì)Gla蛋白(MGP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 基質(zhì)Gla蛋白(MGP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 基質(zhì)Gla蛋白(MGP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 實(shí)驗(yàn)結(jié)果圖
  • SEB477Ra.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過(guò)ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)基質(zhì)Gla蛋白(MGP),經(jīng)檢測(cè)與其它相似物質(zhì)無(wú)明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來(lái)源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)的其它物質(zhì)有交叉反應(yīng)。

回收率

分別于定值血清及血漿樣本中加入一定量的基質(zhì)Gla蛋白(MGP)(加標(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。

樣本 回收率范圍(%) 平均回收率(%)
serum(n=5) 89-97 93
EDTA plasma(n=5) 84-103 98
heparin plasma(n=5) 88-97 93

精密度

精密度用樣品測(cè)定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)算不同濃度樣本的平均值及SD值。
批內(nèi)差: CV<10%
批間差: CV<12%

線性

在定值血清及血漿樣本內(nèi)加入適量的基質(zhì)Gla蛋白(MGP),并倍比稀釋成1:2,1:4,1:8,1:16的待測(cè)樣本,線性范圍即為稀釋后樣本中基質(zhì)Gla蛋白(MGP)含量的測(cè)定值與理論值的比率。

樣本 1:2 1:4 1:8 1:16
serum(n=5) 81-95% 96-103% 78-96% 91-104%
EDTA plasma(n=5) 88-101% 98-105% 78-98% 84-101%
heparin plasma(n=5) 89-96% 89-98% 85-96% 82-93%

穩(wěn)定性

經(jīng)測(cè)定,試劑盒在有效期內(nèi)按推薦溫度保存,其活性降低率小于5%。
為減小外部因素對(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來(lái)進(jìn)行操作可減少人為誤差。

實(shí)驗(yàn)流程

1. 實(shí)驗(yàn)前標(biāo)準(zhǔn)品、試劑及樣本的準(zhǔn)備;
2. 加樣(標(biāo)準(zhǔn)品及樣本)100µL,37°C孵育1小時(shí);
3. 吸棄,加檢測(cè)溶液A100µL,37°C孵育1小時(shí);
4. 洗板3次;
5. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
6. 洗板5次;
7. 加TMB底物90µL,37°C孵育10-20分鐘;
8. 加終止液50µL,立即450nm讀數(shù)。

實(shí)驗(yàn)原理

將基質(zhì)Gla蛋白(MGP)抗體包被于96孔微孔板中,制成固相載體,向微孔中分別加入標(biāo)準(zhǔn)品或標(biāo)本,其中的基質(zhì)Gla蛋白(MGP)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的基質(zhì)Gla蛋白(MGP)抗體,將未結(jié)合的生物素化抗體洗凈后,加入HRP標(biāo)記的親和素,再次徹底洗滌后加入TMB底物顯色。TMB在過(guò)氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成最終的黃色。顏色的深淺和樣品中的基質(zhì)Gla蛋白(MGP)呈正相關(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。

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參考文獻(xiàn)

雜志 參考文獻(xiàn)
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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