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載脂蛋白E(APOE)檢測試劑盒(酶聯(lián)免疫吸附試驗法)

ELISA Kit for Apolipoprotein E (APOE)

Apo-E; AD2; Apoprotein; Alzheimer Disease 2(E4-Associated,Late Onset

  • 載脂蛋白E(APOE)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 產(chǎn)品包裝(模擬)
  • 載脂蛋白E(APOE)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 產(chǎn)品包裝(模擬)
  • 載脂蛋白E(APOE)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 實驗結(jié)果圖
  • SEA704Ra.jpg 標準曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認證

特異性

本試劑盒用于檢測載脂蛋白E(APOE),經(jīng)檢測與其它相似物質(zhì)無明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來源的限制,不可能完成對所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測,因此本試劑盒有可能與未經(jīng)檢測的其它物質(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載脂蛋白E(APOE)(加標樣品),重復(fù)測定并計算其均值,回收率為測定值與理論值的比率。

樣本 回收率范圍(%) 平均回收率(%)
serum(n=5) 94-101 98
EDTA plasma(n=5) 98-105 101
heparin plasma(n=5) 80-95 84

精密度

精密度用樣品測定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對低、中、高值定值樣本進行定量檢測,每份樣本連續(xù)測定20 次,分別計算不同濃度樣本的平均值及SD值。
批間差:選取3個不同批次的試劑盒分別對低、中、高值定值樣本進行定量測定,每個樣本使用同一試劑盒重復(fù)測定8次,分別計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)加入適量的載脂蛋白E(APOE),并倍比稀釋成1:2,1:4,1:8,1:16的待測樣本,線性范圍即為稀釋后樣本中載脂蛋白E(APOE)含量的測定值與理論值的比率。

樣本 1:2 1:4 1:8 1:16
serum(n=5) 92-101% 84-101% 87-95% 93-101%
EDTA plasma(n=5) 81-89% 79-101% 93-104% 92-101%
heparin plasma(n=5) 98-105% 91-102% 88-97% 79-95%

穩(wěn)定性

經(jīng)測定,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%。
為減小外部因素對試劑盒破壞前后檢測值的影響,實驗室的環(huán)境條件需盡量保持一致,尤其是實驗室內(nèi)溫度、濕度及溫育條件。其次由同一實驗員來進行操作可減少人為誤差。

實驗流程

1. 實驗前標準品、試劑及樣本的準備;
2. 加樣(標準品及樣本)100µL,37°C孵育1小時;
3. 吸棄,加檢測溶液A100µL,37°C孵育1小時;
4. 洗板3次;
5. 加檢測溶液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)。

實驗原理

將載脂蛋白E(APOE)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準品或標本,其中的載脂蛋白E(APOE)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的載脂蛋白E(APOE)抗體,將未結(jié)合的生物素化抗體洗凈后,加入HRP標記的親和素,再次徹底洗滌后加入TMB底物顯色。TMB在過氧化物酶的催化下轉(zhuǎn)化成藍色,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載脂蛋白E(APOE)呈正相關(guān)。用酶標儀在450nm波長下測定吸光度(O.D.值),計算樣品濃度。

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參考文獻

雜志 參考文獻
Seizure - European Journal of Epilepsy Cerebrospinal fluid apolipoprotein E concentration decreases after seizure [Seizure: PIIS1059131109002441]
Biochimica et Biophysica Acta Hepatic lipase- and endothelial lipase-deficiency in mice promotes macrophage-to-feces RCT and HDL antioxidant properties [PubMed: 23328279 ]
Medical Oncology Expression and clinical significance of apolipoprotein E in pancreatic ductal adenocarcinoma [Pubmed: 23609192]
Diabetes. Adipocyte Atp-Binding Cassette G1 Promotes Triglyceride Storage, Fat Mass Growth And Human Obesity [Pubmed:25249572]
renal failure Frequencies of apolipoprotein E alleles in depressed patients undergoing hemodialysis – a case-control study [PubMed: 25707516]
Biochim Biophys Acta Culturing of HepG2 cells with human serum improve their functionality and suitability in studies of lipid metabolism [PubMed: 26515253]
Diabetes Adipocyte ATP-binding cassette G1 promotes triglyceride storage, fat mass growth, and human obesity [PubMed: 25249572]
Scientific Reports Adaptive?immunity?against?gut?microbiota?enhances?apoE-mediated?immune?regulation?andreduces?atherosclerosis?and?western-diet-related?inflammation. [pubmed:27383250]
Biochemical Pharmacology? Differential effects of apoE and apoJ mimetic peptides on the action of an anti-Aβ scFv in 3xTg-AD mice [Pubmed:30026023]
Oncotarget Deletion of plasma Phospholipid Transfer Protein (PLTP) increases microglial phagocytosis and reduces cerebral amyloid-β deposition in the J20 mouse?… [Pubmed:29731975]
Proteomics Clinical Applications Comparative proteomics analysis of serum proteins in gestational diabetes during early and middle stages of pregnancy [Pubmed: 31162828]
Nature Communications Hepatic HuR modulates lipid homeostasis in response to high-fat diet [Pubmed: 32546794]
VETERINARY PATHOLOGY Pathology and Proteomics-Based Diagnosis of Localized Light-Chain Amyloidosis in Dogs and Cats [Pubmed: 32880234]
Journal of Clinical Medicine The Significance of Apolipoprotein E Measurement in the Screening of Fetal Down Syndrome [33321701]
Cancer Cell Int Discovery and validation of FBLN1 and ANT3 as potential biomarkers for early detection of cervical cancer [33602229]
Nat Commun GP73 is a TBC-domain Rab GTPase-activating protein contributing to the pathogenesis of non-alcoholic fatty liver disease without obesity [34853313]
Talanta Iridium nanoclusters as high sensitive-tunable elemental labels for immunoassays: Determination of IgE and APOE in aqueous humor by inductively coupled?… [Pubmed:35378356]
Analytica Chimica Acta Gold nanoclusters as elemental label for the sequential quantification of apolipoprotein E and metallothionein 2A in individual human cells of the retinal?… [Pubmed:35361429]
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