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堿性磷酸酶(ALP)檢測試劑盒(酶聯(lián)免疫吸附試驗法)

ELISA Kit for Alkaline Phosphatase (ALP)

AKP; ALKP; Basic Phosphatase

  • 堿性磷酸酶(ALP)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 產(chǎn)品包裝(模擬)
  • 堿性磷酸酶(ALP)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 產(chǎn)品包裝(模擬)
  • 堿性磷酸酶(ALP)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 實驗結(jié)果圖
  • SEB472Bo.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

本試劑盒用于檢測堿性磷酸酶(ALP),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堿性磷酸酶(ALP)(加標(biāo)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) 96-105 101
EDTA plasma(n=5) 87-95 91
heparin plasma(n=5) 93-101 97

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)加入適量的堿性磷酸酶(ALP),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堿性磷酸酶(ALP)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) 85-103% 80-101% 81-93% 83-92%
EDTA plasma(n=5) 78-95% 89-96% 79-89% 87-101%
heparin plasma(n=5) 91-101% 92-102% 78-97% 78-95%

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(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;
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

將堿性磷酸酶(ALP)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,其中的堿性磷酸酶(ALP)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的堿性磷酸酶(ALP)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堿性磷酸酶(ALP)呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(O.D.值),計算樣品濃度。

相關(guān)產(chǎn)品

編號 適用物種:Bos taurus; Bovine (Cattle,牛) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
RPB472Bo01 堿性磷酸酶(ALP)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
PAB472Bo01 堿性磷酸酶(ALP)多克隆抗體 WB; IHC; ICC; IP.
LAB472Bo71 堿性磷酸酶(ALP)多克隆抗體(生物素標(biāo)記) WB; IHC; ICC.
SEB472Bo 堿性磷酸酶(ALP)檢測試劑盒(酶聯(lián)免疫吸附試驗法) Enzyme-linked immunosorbent assay for Antigen Detection.
SCB472Bo 堿性磷酸酶(ALP)檢測試劑盒(化學(xué)發(fā)光免疫分析法) Chemiluminescent immunoassay for Antigen Detection.
LMB472Bo 堿性磷酸酶(ALP)等多因子檢測試劑盒(流式熒光發(fā)光法) FLIA Kit for Antigen Detection.
KSB472Bo01 堿性磷酸酶(ALP)檢測試劑盒DIY材料(酶聯(lián)免疫吸附試驗法) Main materials for "Do It (ELISA Kit) Yourself".

參考文獻

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