齊一生物鞘磷脂（SM）試劑盒

齊一生物鞘磷脂（SM）試劑盒齊一生物長(zhǎng)期經(jīng)營(yíng)ELISA試劑盒（免費(fèi)代測(cè)）及抗體,分子學(xué)生物試劑盒,金標(biāo)法檢測(cè)試劑盒,比色法檢測(cè)試劑盒,細(xì)胞菌株,培養(yǎng)基,細(xì)胞因子,生化試劑,耗材等生物試劑產(chǎn)品.誠(chéng)信經(jīng)營(yíng)，價(jià)格實(shí)惠，服務(wù)周到，質(zhì)量保障！提供各種實(shí)驗(yàn)代測(cè)服務(wù).歡迎新老客戶(hù)！齊一生物科技（上海）有限公司，專(zhuān)業(yè)生產(chǎn)研發(fā)生化科研試劑全國(guó)：

齊一生物鞘磷脂（SM）試劑盒1.包被過(guò)程(注意設(shè)置空白對(duì)照,陰性對(duì)照):
將所用抗原用包被稀釋液稀釋到適當(dāng)濃度(一般所需抗原包被量為每孔20-200μg),每孔抗原加入100μl,置37℃,4h,或4℃,24h;棄去孔中液體.(為避免蒸發(fā),板上應(yīng)加蓋或?qū)迤椒旁诘撞坑袧窦啿嫉慕饘贊窈兄?
2.封閉酶標(biāo)反應(yīng)孔:
5%小牛血清置37℃封閉40min.封閉時(shí)將封閉液加滿(mǎn)各反應(yīng)孔,并去除各孔中的氣泡,封閉結(jié)束后用洗滌液滿(mǎn)孔洗滌3遍,每遍3min.
洗滌方法:吸干孔內(nèi)反應(yīng)液,將洗滌液注滿(mǎn)板孔,放置2min略作搖動(dòng),吸干孔內(nèi)液,傾去液體后在吸水紙上拍干
洗滌次數(shù)3次
3.加入待檢測(cè)樣品(建立合適的濃度梯度):
檢測(cè)時(shí)一般采用1:50-1:400的稀釋度,應(yīng)采用較大稀釋體積進(jìn)行,一般保證樣品吸取量>20μl.
將稀釋好的樣品加入酶標(biāo)反應(yīng)孔中,每樣品少加雙孔,每孔100μl,置于37℃,40-60min.用洗滌液滿(mǎn)孔洗滌3遍,每遍3min.
4.加入酶標(biāo)抗體:
酶標(biāo)抗體:根據(jù)酶結(jié)合物提供商提供的參考工作稀釋度進(jìn)行.37℃,30-60min之間.短于30min往往結(jié)果不穩(wěn)定.每孔加100μl.洗滌同前.
5.加入底物液(現(xiàn)用現(xiàn)配):
*TMB-過(guò)氧化氫尿素溶液,OPD-過(guò)氧化氫底物液系統(tǒng)次之.
底物加入量:每孔100μl,置37℃避光放置3-5分鐘,加入終止液顯色.
6.終止反應(yīng):
每孔加入終止液50μl終止反應(yīng),于20min內(nèi)測(cè)定實(shí)驗(yàn)結(jié)果.
7.結(jié)果判斷:
OPD顯色后采用492nm波長(zhǎng),TMB反應(yīng)產(chǎn)物檢測(cè)需要450nm波長(zhǎng).檢測(cè)時(shí)一定要首先進(jìn)行空白孔系統(tǒng)調(diào)零.用測(cè)定標(biāo)本孔的吸收值與一組陰性標(biāo)本測(cè)定孔平均值的比值(P/N)表示,當(dāng)P/N大于2時(shí)作為抗體的效價(jià).(數(shù)值的大小依具體檢測(cè)要求而定.)

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