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Ⅳ型膠原(COL4)檢測試劑盒(酶聯(lián)免疫吸附試驗法)

ELISA Kit for Collagen Type IV (COL4)

Type-IV Collagen

  • Ⅳ型膠原(COL4)檢測試劑盒(酶聯(lián)免疫吸附試驗法)產(chǎn)品包裝(模擬)
  • Ⅳ型膠原(COL4)檢測試劑盒(酶聯(lián)免疫吸附試驗法)產(chǎn)品包裝(模擬)
  • Ⅳ型膠原(COL4)檢測試劑盒(酶聯(lián)免疫吸附試驗法)實驗結(jié)果圖
  • SEA180Rb.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Ⅳ型膠原(COL4),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Ⅳ型膠原(COL4)(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)88-9692
EDTA plasma(n=5)83-9387
heparin plasma(n=5)88-10193

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Ⅳ型膠原(COL4),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Ⅳ型膠原(COL4)含量的測定值與理論值的比率。

樣本1:21:41:81:16
serum(n=5)80-92%90-102%93-101%87-102%
EDTA plasma(n=5)98-105%83-99%80-99%86-97%
heparin plasma(n=5)91-101%81-101%85-95%95-104%

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Ⅳ型膠原(COL4)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Ⅳ型膠原(COL4)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的Ⅳ型膠原(COL4)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Ⅳ型膠原(COL4)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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SEA180RbⅣ型膠原(COL4)檢測試劑盒(酶聯(lián)免疫吸附試驗法)Enzyme-linked immunosorbent assay for Antigen Detection.
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參考文獻

雜志參考文獻
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