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Ⅳ型膠原(COL4)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Collagen Type IV (COL4)

Type-IV Collagen

  • Ⅳ型膠原(COL4)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)產(chǎn)品包裝(模擬)
  • Ⅳ型膠原(COL4)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)產(chǎn)品包裝(模擬)
  • Ⅳ型膠原(COL4)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
  • SEA180Hu.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)Ⅳ型膠原(COL4),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Ⅳ型膠原(COL4)(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)85-9992
EDTA plasma(n=5)92-9995
heparin plasma(n=5)92-10196

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Ⅳ型膠原(COL4),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Ⅳ型膠原(COL4)含量的測(cè)定值與理論值的比率。

樣本1:21:41:81:16
serum(n=5)96-105%94-105%90-99%86-98%
EDTA plasma(n=5)79-92%79-104%84-98%89-101%
heparin plasma(n=5)79-104%94-103%81-98%80-103%

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(biāo)準(zhǔn)品或標(biāo)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(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Ⅳ型膠原(COL4)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。

相關(guān)產(chǎn)品

編號(hào)適用物種:Homo sapiens (Human,人)應(yīng)用(僅供研究使用,不用于臨床診斷!)
NPA180Hu01Ⅳ型膠原(COL4)天然蛋白Positive Control; Immunogen; SDS-PAGE; WB.
PAA180Hu01Ⅳ型膠原(COL4)多克隆抗體WB; IHC; ICC; IP.
LAA180Hu71Ⅳ型膠原(COL4)多克隆抗體(生物素標(biāo)記)WB; IHC; ICC.
SEA180HuⅣ型膠原(COL4)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)Enzyme-linked immunosorbent assay for Antigen Detection.
HEA180HuⅣ型膠原(COL4)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,高敏型)Enzyme-linked immunosorbent assay for Antigen Detection.
SCA180HuⅣ型膠原(COL4)檢測(cè)試劑盒(化學(xué)發(fā)光免疫分析法)Chemiluminescent immunoassay for Antigen Detection.
LMA180HuⅣ型膠原(COL4)等多因子檢測(cè)試劑盒(流式熒光發(fā)光法)FLIA Kit for Antigen Detection.
KSA180Hu01Ⅳ型膠原(COL4)檢測(cè)試劑盒DIY材料(酶聯(lián)免疫吸附試驗(yàn)法)Main materials for "Do It (ELISA Kit) Yourself".

參考文獻(xiàn)

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