紅細胞生成素(EPO)檢測試劑盒(酶聯(lián)免疫吸附試驗法)
編號SEA028Mu
物種Mus musculus (Mouse，小鼠) 相同的名稱，不同的物種。
實驗方法雙抗夾心
反應時長3h
檢測范圍6.25-400pg/mL
靈敏度最小可檢測劑量小于等于2.68pg/mL.
樣本類型serum, plasma, tissue homogenates, cell lysates and other biological fluids
下載英文說明書 中文說明書
規(guī)格48T96T 96T*5 96T*10 96T*100
價格￥ 2988
特異性
本試劑盒用于檢測紅細胞生成素(EPO)，經(jīng)檢測與其它相似物質(zhì)無明顯交叉反應。
由于受到技術及樣本來源的限制，不可能完成對所有相關或相似物質(zhì)交叉反應檢測，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。
回收率
分別于定值血清及血漿樣本中加入一定量的紅細胞生成素(EPO)（加標樣品），重復測定并計算其均值，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) | 84-102 | 89 |
| EDTA plasma(n=5) | 92-101 | 95 |
| heparin plasma(n=5) | 88-105 | 95 |
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)加入適量的紅細胞生成素(EPO)，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紅細胞生成素(EPO)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) | 89-97% | 91-99% | 96-105% | 78-94% |
| EDTA plasma(n=5) | 82-102% | 92-99% | 82-91% | 86-97% |
| heparin plasma(n=5) | 99-105% | 85-97% | 81-101% | 86-94% |
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
將紅細胞生成素(EPO)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，其中的紅細胞生成素(EPO)與連接于固相載體上的抗體結合，然后加入生物su化的紅細胞生成素(EPO)抗體，將未結合的生物su化抗體洗凈后，加入HRP標記的親和素，再次洗滌后加入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紅細胞生成素(EPO)呈正相關。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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