膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)
ELISA Kit for Glial Cell Line Derived Neurotrophic Factor (GDNF)
ATF1; ATF2; HFB1-GDNF; Astrocyte-Derived Trophic Factor; Glial Derived Neurotrophic Factor
- 編號SEA043Mu
- 物種Mus musculus (Mouse,小鼠) 相同的名稱,不同的物種。
- 實(shí)驗(yàn)方法雙抗夾心
- 反應(yīng)時長3h
- 檢測范圍78-5,000pg/mL
- 靈敏度最小可檢測劑量小于等于32pg/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
- 價格 ¥ 2722 ¥ 3888 ¥ 17496 ¥ 33048 ¥ 272160
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特異性
本試劑盒用于檢測膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF),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膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF)(加標(biāo)樣品),重復(fù)測定并計(jì)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) | 78-96 | 89 |
EDTA plasma(n=5) | 85-99 | 93 |
heparin plasma(n=5) | 89-97 | 92 |
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(jì)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(jì)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)加入適量的膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF),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膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF)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) | 91-99% | 82-89% | 81-103% | 93-102% |
EDTA plasma(n=5) | 96-104% | 84-92% | 91-105% | 95-105% |
heparin plasma(n=5) | 83-102% | 96-104% | 99-105% | 78-103% |
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(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來進(jìn)行操作可減少人為誤差。
實(shí)驗(yàn)流程
1. 實(shí)驗(yàn)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(shí)驗(yàn)原理
將膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF)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,其中的膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF)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膠質(zhì)細(xì)胞系來源神經(jīng)營養(yǎng)因子(GDNF)呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(O.D.值),計(jì)算樣品濃度。
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參考文獻(xiàn)
雜志 | 參考文獻(xiàn) |
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