操作步驟

1. 標(biao)(biao)準(zhun)品(pin)(pin)的稀(xi)(xi)釋(shi)(shi)(shi)與(yu)加(jia)(jia)樣:在酶標(biao)(biao)包(bao)被板上設標(biao)(biao)準(zhun)品(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)10孔(kong)(kong)(kong)(kong)(kong)，在一、 二(er)(er)(er)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)加(jia)(jia)標(biao)(biao)準(zhun)品(pin)(pin)100ul，然后(hou)(hou)在一、二(er)(er)(er)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)加(jia)(jia)標(biao)(biao)準(zhun)品(pin)(pin)稀(xi)(xi)釋(shi)(shi)(shi)液(ye)50ul，混(hun)勻(yun)(yun); 然后(hou)(hou)從(cong)一孔(kong)(kong)(kong)(kong)(kong)、二(er)(er)(er)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)各(ge)(ge)取100ul分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)加(jia)(jia)到(dao)第(di)(di)(di)三(san)孔(kong)(kong)(kong)(kong)(kong)和(he)第(di)(di)(di)四(si)孔(kong)(kong)(kong)(kong)(kong)，再(zai)在第(di)(di)(di)三(san)、第(di)(di)(di)四(si)孔(kong)(kong)(kong)(kong)(kong)分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)加(jia)(jia)標(biao)(biao)準(zhun)品(pin)(pin)稀(xi)(xi)釋(shi)(shi)(shi)液(ye)50ul，混(hun)勻(yun)(yun);然后(hou)(hou)在第(di)(di)(di)三(san)孔(kong)(kong)(kong)(kong)(kong)和(he)第(di)(di)(di)四(si)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)先各(ge)(ge)取50ul棄掉，再(zai)各(ge)(ge)取50ul分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)加(jia)(jia)到(dao)第(di)(di)(di)五、第(di)(di)(di)六(liu)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)，再(zai)在第(di)(di)(di)五、第(di)(di)(di)六(liu)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)加(jia)(jia)標(biao)(biao)準(zhun)品(pin)(pin)稀(xi)(xi)釋(shi)(shi)(shi)液(ye)50ul，混(hun)勻(yun)(yun);混(hun)勻(yun)(yun)后(hou)(hou)從(cong)第(di)(di)(di)五、第(di)(di)(di)六(liu)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)各(ge)(ge)取50ul分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)加(jia)(jia)到(dao)第(di)(di)(di)七、第(di)(di)(di)八(ba)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)，再(zai)在第(di)(di)(di)七、第(di)(di)(di)八(ba)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)加(jia)(jia)標(biao)(biao)準(zhun)品(pin)(pin)稀(xi)(xi)釋(shi)(shi)(shi)液(ye)50u，混(hun)勻(yun)(yun)后(hou)(hou)從(cong)第(di)(di)(di)七、第(di)(di)(di)八(ba)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)取50ul加(jia)(jia)到(dao)第(di)(di)(di)九、第(di)(di)(di)十孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)，再(zai)在第(di)(di)(di)九第(di)(di)(di)十孔(kong)(kong)(kong)(kong)(kong)分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)加(jia)(jia)標(biao)(biao)準(zhun)品(pin)(pin)稀(xi)(xi)釋(shi)(shi)(shi)液(ye)50μ，混(hun)勻(yun)(yun)后(hou)(hou)從(cong)第(di)(di)(di)九第(di)(di)(di)十孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)各(ge)(ge)取50μl棄掉。(稀(xi)(xi)釋(shi)(shi)(shi)后(hou)(hou)各(ge)(ge)孔(kong)(kong)(kong)(kong)(kong)加(jia)(jia)樣量(liang)都為50μl，濃度分(fen)(fen)(fen)(fen)別(bie)(bie)(bie)為240pg/ml，160pg/ml，80pg/ml，40pg/ml，20pg/ml)。

2. 加樣(yang)(yang): 分(fen)別設(she)空白(bai)孔(空白(bai)對照孔不(bu)加樣(yang)(yang)品(pin)及酶(mei)標(biao)試劑(ji)，其余(yu)各步操作相同)、待(dai)測(ce)樣(yang)(yang)品(pin)孔。在酶(mei)標(biao)包被板(ban)上待(dai)測(ce)樣(yang)(yang)品(pin)孔中先(xian)加樣(yang)(yang)品(pin)稀釋液40ul，然后再加待(dai)測(ce)樣(yang)(yang)品(pin)10ul(樣(yang)(yang)品(pin)zui終(zhong)稀釋度為(wei)5倍)。加樣(yang)(yang)將樣(yang)(yang)品(pin)加于酶(mei)標(biao)板(ban)孔底部，盡量不(bu)觸(chu)及孔壁(bi)，輕輕晃動混勻(yun)。

3. 溫育: 用(yong)封板(ban)膜封板(ban)后(hou)置(zhi)37°℃溫育30分鐘。

4. 配(pei)液: 將20倍(bei)濃縮洗滌(di)液用(yong)蒸餾水20倍(bei)稀釋后(hou)備用(yong)。

5. 洗滌: 小心(xin)揭掉(diao)封板膜，棄去液體，甩干(gan)，每孔加滿洗滌液，靜(jing)置30秒后棄去，如(ru)此重復5次，拍干(gan)。

6. 加酶: 每孔加入(ru)酶標試劑50ul，空白孔除(chu)外。

7. 溫育(yu): 操作同3。

8. 洗滌: 操作同5。

9. 顯色(se): 每孔先加(jia)(jia)入顯色(se)劑A50ul，再加(jia)(jia)入顯色(se)劑B50ul，輕輕震蕩混勻(yun)，37°C避光顯色(se)15分鐘.以(yi)內(nei)進行。

10. 終(zhong)止: 每孔加(jia)終(zhong)止液50μl，終(zhong)止反應(ying)(此時藍色(se)立轉黃色(se))。

11. 測定: 以(yi)空白空調零，450nm波長依(yi)序測量各孔的吸光度(du)(OD值)。測定應在(zai)加(jia)終止液后(hou)15分鐘。

上海研域生物科技有限公司成立于上海浦東新區。是一家專門從事人ELISA試劑盒生(sheng)(sheng)物(wu)技(ji)術相關(guan)產(chan)品(pin)，勵(li)志研發(fa)和(he)銷售的(de)(de)綜合性(xing)生(sheng)(sheng)物(wu)公司，產(chan)品(pin)遠銷多個國(guo)家和(he)地(di)區(qu)，我們將一(yi)(yi)如既往地(di)秉承(cheng)“一(yi)(yi)切為(wei)了滿足(zu)客戶的(de)(de)需求"的(de)(de)經(jing)營(ying)宗旨; 牢固樹(shu)立“以(yi)客戶需求為(wei)準則、以(yi)市場需求為(wei)導向，不斷開發(fa)高質量的(de)(de)新產(chan)品(pin)，提供客戶滿意的(de)(de)綜合服務質量"的(de)(de)方(fang)針。