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磷脂酰肌醇蛋白聚糖3(GPC3)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Glypican 3 (GPC3)

DGSX; OCI5; SDYS; SGB; SGBS1; MXR7; Glypican Proteoglycan 3; GTR2-2; Intestinal protein OCI-5; Secreted glypican-3

  • 磷脂酰肌醇蛋白聚糖3(GPC3)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 磷脂酰肌醇蛋白聚糖3(GPC3)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 磷脂酰肌醇蛋白聚糖3(GPC3)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
  • SEA971Hu.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)磷脂酰肌醇蛋白聚糖3(GPC3),經(jīng)檢測(cè)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(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磷脂酰肌醇蛋白聚糖3(GPC3)(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) 78-99 95
EDTA plasma(n=5) 90-104 94
heparin plasma(n=5) 98-105 101

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)加入適量的磷脂酰肌醇蛋白聚糖3(GPC3),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磷脂酰肌醇蛋白聚糖3(GPC3)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) 79-101% 78-94% 82-89% 88-98%
EDTA plasma(n=5) 88-101% 78-94% 94-105% 78-96%
heparin plasma(n=5) 93-101% 90-104% 87-94% 87-97%

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

將磷脂酰肌醇蛋白聚糖3(GPC3)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,其中的磷脂酰肌醇蛋白聚糖3(GPC3)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的磷脂酰肌醇蛋白聚糖3(GPC3)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磷脂酰肌醇蛋白聚糖3(GPC3)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)
Nature and Science Diagnostic Role of Serum Glypican-3 as a Tumor Marker for Hepatocellular Carcinoma [Sciencepub:10043238]
Journal of Applied Pharmaceutical Science Cytotoxic effects of antiglypican-3 against HepG2 cell line [Ebscohost: Source]
Asian Pacific Journal of Cancer Prevention Can Glypican3 be Diagnostic for Early Hepatocellular Carcinoma among Egyptian Patients? [Pubmed: 24460300]
Clin Biochem. Diagnostic accuracy of serum glypican-3 for hepatocellular carcinoma: A systematic review and meta-analysis [Pubmed: 24362268]
Science and Nature Diagnostic Role of Serum Glypican-3 as Hepatocellular Carcinoma [Zolcat: Source]
Alexandria Journal of Medicine Role of serum glypican-3 in the diagnosis and differentiation of small hepatocellular carcinoma from hepatitis-C virus cirrhosis [ScienceDirect: S2090506814000062]
J Egypt Natl Canc Inst. Comparing prothrombin induced by vitamin K absence-II (PIVKA-II) with the oncofetal proteins glypican-3, Alpha feto protein and carcinoembryonic antigen in diagnosing hepatocellular carcinoma among Egyptian patients. [Pubmed:24841158]
European Journal of Pharmacology Evaluation of antiglypican-3 therapy as a promising target for amelioration of hepatic tissue damage in hepatocellular carcinoma [Pubmed:25449037]
Hepatobiliary Pancreat Dis Int Predictive value of tumor markers in patients with recurrent hepatocellular carcinoma in different vascular invasion pattern [abstract:abstract4525.shtml]
Clinical?and Molecular Hepatology? Glypican-3 level assessed by the enzyme-linked immunosorbent assay is inferior to alpha-fetoprotein level for hepatocellular carcinoma diagnosis [pmc:articles]
Wiley Glypican‐3: A promising biomarker for hepatocellular carcinoma diagnosis and treatment [pubmed:28621802]
Biomedicine & Pharmacotherapy Cytotoxic and partial hepatoprotective activity of sodium ascorbate against hepatocellular carcinoma through inhibition of sulfatase-2 and [Pubmed:29669302]
Journal of Cancer and Clinical Oncology Development and Validation of a Two-Site Immunoradiometric assay for Glypican-3 in plasma: Implication in Diagnosis of Hepatocellular Carcinoma [Researchgate:Source]
Turkish?journal?of?oncology Serum Glypican-3, Vascular Endothelial Growth Factor, and Interleukin-6 Levels in Hepatocellular Carcinoma [10.5505:tjo.2018.1742]
Intensive?Care?Medicine?Experimental Elevated plasma glypicans are associated with organ failure in patients with infection [Pubmed: 30618011]
VALUE OF SERUM GLYPICAN 3 AS A PREDICTIVE TOOL FOR EARLY DIAGNOSIS OF HEPATOCELLULAR CARCINOMA (HCC) []
ADVANCED RESEARCHES Search for Effective Serum Tumor Markers for Early Diagnosis of Hepatocellular Carcinoma Associated with Hepatitis C []
Life Sci Selenium nanoparticles overcomes sorafenib resistance in thioacetamide induced hepatocellular carcinoma in rats by modulation of mTOR, NF-κB pathways and?… [Pubmed:35640776]
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