促腎上腺皮質(zhì)激素(ACTH)檢測試劑盒(酶聯(lián)免疫吸附試驗法)
ELISA Kit for Adrenocorticotropic Hormone (ACTH)
Corticotropin; Adrenocorticotropin
- 編號CEA836Cp
- 物種Capra hircus; Caprine (Goat,山羊) 相同的名稱,不同的物種。
- 實驗方法競爭抑制
- 反應(yīng)時長2h
- 檢測范圍12.35-1000pg/mL
- 靈敏度最小可檢測劑量小于等于4.92pg/mL.
- 樣本類型Serum, plasma, tissue homogenates, cell lysates, cell culture supernates 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
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特異性
本試劑盒用于檢測促腎上腺皮質(zhì)激素(ACTH),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促腎上腺皮質(zhì)激素(ACTH)(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) | 92-99 | 95 |
EDTA plasma(n=5) | 79-91 | 85 |
heparin plasma(n=5) | 83-93 | 89 |
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)加入適量的促腎上腺皮質(zhì)激素(ACTH),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促腎上腺皮質(zhì)激素(ACTH)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) | 94-104% | 96-105% | 98-105% | 95-103% |
EDTA plasma(n=5) | 85-93% | 83-104% | 80-99% | 78-89% |
heparin plasma(n=5) | 84-92% | 89-101% | 80-91% | 81-91% |
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)50µL,
加入50µL檢測液A(臨用前配制);
37°C溫育1小時。
3. 洗板3次;
4. 加檢測溶液B100µL,37°C孵育30分鐘;
5. 洗板5次;
6. 加TMB底物90µL,37°C孵育10-20分鐘;
7. 加終止液50µL,立即450nm讀數(shù)。
實驗原理
本試劑盒應(yīng)用競爭抑制酶聯(lián)免疫分析法測定標(biāo)本中待測物質(zhì)水平。將促腎上腺皮質(zhì)激素(ACTH)單克隆抗體包被微孔板,制成固相載 體,往包被抗體的微孔中同時加入生物素標(biāo)記的抗原和待測抗原(標(biāo)準(zhǔn)品或樣本),待測抗原與生物素標(biāo)記抗原對特異性抗體進(jìn)行競爭結(jié)合。溫育后經(jīng)洗滌去掉未結(jié)合物,然后加入HRP標(biāo)記的親和素,經(jīng)過溫育和徹底洗滌后加入底物TMB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。待測標(biāo)本濃度越高,標(biāo)記抗原和抗體的結(jié)合就越受到抑制,顯色愈淺。顯色的深淺與酶量呈正相關(guān),而與樣品中待測物質(zhì)含量呈負(fù)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:Capra hircus; Caprine (Goat,山羊) | 應(yīng)用(僅供研究使用,不用于臨床診斷!) |
CEA836Cp | 促腎上腺皮質(zhì)激素(ACTH)檢測試劑盒(酶聯(lián)免疫吸附試驗法) | Enzyme-linked immunosorbent assay for Antigen Detection. |
LMA836Cp | 促腎上腺皮質(zhì)激素(ACTH)等多因子檢測試劑盒(流式熒光發(fā)光法) | FLIA Kit for Antigen Detection. |
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