血管內皮生長因子A(VEGFA)檢測試劑盒(酶聯免疫吸附試驗法)
編號SEA143Ca
物種Canis familiaris; Canine (Dog，犬) 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15.6-1,000pg/mL
靈敏度最小可檢測劑量小于等于5.6pg/mL.
樣本類型Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
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規格48T96T 96T*5 96T*10 96T*100
價格￥ 2008
產品包裝圖模擬
特異性
本試劑盒用于檢測血管內皮生長因子A(VEGFA)，經檢測與其它相似物質無明顯交叉反應。
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交叉反應檢測，因此本試劑盒有可能與未經檢測的其它物質有交叉反應。
回收率
分別于定值血清及血漿樣本中加入一定量的血管內皮生長因子A(VEGFA)（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) | 88-95 | 92 |
| EDTA plasma(n=5) | 83-93 | 88 |
| heparin plasma(n=5) | 88-102 | 91 |
精密度
精密度用樣品測定值的變異系數CV表示。CV(%) = SD/mean×100
批內差：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。
批內差： CV<10%
批間差： CV<12%
線性
在定值血清及血漿樣本內加入適量的血管內皮生長因子A(VEGFA)，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血管內皮生長因子A(VEGFA)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) | 79-102% | 79-103% | 94-103% | 79-97% |
| EDTA plasma(n=5) | 89-98% | 93-104% | 90-98% | 83-98% |
| heparin plasma(n=5) | 84-91% | 78-88% | 78-98% | 87-105% |
穩定性
經測定，試劑盒在有效期內按推薦溫度保存，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，實驗室的環境條件需盡量保持一致，尤其是實驗室內溫度、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。
實驗原理
將血管內皮生長因子A(VEGFA)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，其中的血管內皮生長因子A(VEGFA)與連接于固相載體上的抗體結合，然后加入生物-素化的血管內皮生長因子A(VEGFA)抗體，將未結合的生物-素化抗體洗凈后，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化成藍色，并在酸的作用下轉化成最終的黃色。顏色的深淺和樣品中的血管內皮生長因子A(VEGFA)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。
參考文獻
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