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饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本)

Mini Samples ELISA Kit for Ghrelin (GHRL)

MTLRP; Growth Hormone-Releasing Peptide

  • 饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本) 產(chǎn)品包裝(模擬)
  • 饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本) 產(chǎn)品包裝(模擬)
  • 饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本) 實(shí)驗(yàn)結(jié)果圖
  • MEA991Ra.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)饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本),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饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本)(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) 98-105 101
EDTA plasma(n=5) 81-97 88
heparin plasma(n=5) 78-104 97

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)加入適量的饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本),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饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本)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) 87-101% 97-105% 86-99% 78-88%
EDTA plasma(n=5) 78-104% 85-92% 78-91% 93-101%
heparin plasma(n=5) 82-101% 80-98% 88-95% 81-101%

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)25µL后,立即加入檢測(cè)溶液A25µL,
    37°C孵育1小時(shí);
3. 甩干,洗板3次;
4. 加檢測(cè)溶液B50µL,37°C孵育30分鐘;
5. 洗板5次;
6. 加TMB底物50µL,37°C孵育10-20分鐘;
7. 加終止液25µL,立即450nm讀數(shù)。

實(shí)驗(yàn)原理

本試劑盒應(yīng)用競(jìng)爭(zhēng)抑制酶聯(lián)免疫分析法測(cè)定標(biāo)本中待測(cè)物質(zhì)水平。將饑餓素(GHRL)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本)單克隆抗體包被微孔板,制成固相載體,往包被抗體的微孔中同時(shí)加入生物素標(biāo)記的抗原和待測(cè)抗原(標(biāo)準(zhǔn)品或樣本),待測(cè)抗原與生物素標(biāo)記抗原對(duì)特異性抗體進(jìn)行競(jìng)爭(zhēng)結(jié)合。溫育后經(jīng)洗滌去掉未結(jié)合物,然后加入HRP標(biāo)記的親和素,經(jīng)過溫育和徹底洗滌后加入底物TMB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。待測(cè)標(biāo)本濃度越高,標(biāo)記抗原和抗體的結(jié)合就越受到抑制,顯色愈淺。顯色的深淺與酶量呈正相關(guān),而與樣品中待測(cè)物質(zhì)含量呈負(fù)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)
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