視黃-醇結(jié)合蛋白4(RBP4)檢測試劑盒(酶聯(lián)免疫吸附試驗法)
編號SEA929Hu
物種Homo sapiens (Human，人) 相同的名稱，不同的物種。
實驗方法雙抗夾心
反應(yīng)時長3h
檢測范圍0.625-40ng/mL
靈敏度最小可檢測劑量小于等于0.267ng/mL.
樣本類型Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
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規(guī)格48T96T 96T*5 96T*10 96T*100
價格￥ 2092
產(chǎn)品包裝圖模擬
特異性
本試劑盒用于檢測視-黃醇結(jié)合蛋白4(RBP4)，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視-黃醇結(jié)合蛋白4(RBP4)（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) | 96-103 | 99 |
| EDTA plasma(n=5) | 84-99 | 90 |
| heparin plasma(n=5) | 92-101 | 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(jìn)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(jìn)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)加入適量的視-黃醇結(jié)合蛋白4(RBP4)，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視-黃醇結(jié)合蛋白4(RBP4)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) | 92-101% | 83-104% | 96-104% | 96-105% |
| EDTA plasma(n=5) | 80-102% | 90-98% | 81-97% | 87-94% |
| heparin plasma(n=5) | 84-92% | 95-105% | 78-91% | 92-99% |
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(jìn)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
將視-黃醇結(jié)合蛋白4(RBP4)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，其中的視-黃醇結(jié)合蛋白4(RBP4)與連接于固相載體上的抗體結(jié)合，然后加入生物-素化的視-黃醇結(jié)合蛋白4(RBP4)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視-黃醇結(jié)合蛋白4(RBP4)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。
參考文獻(xiàn)
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