1. 標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)的稀(xi)釋(shi)(shi)(shi)(shi)與加(jia)(jia)(jia)樣：羊ELISA試劑盒(he)在(zai)酶(mei)標(biao)(biao)(biao)包被板上(shang)設標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)孔(kong)(kong)10孔(kong)(kong)，在(zai)第(di)(di)(di)(di)(di)(di)一、第(di)(di)(di)(di)(di)(di)二(er)孔(kong)(kong)中(zhong)分(fen)(fen)別(bie)加(jia)(jia)(jia)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)100μl，然(ran)(ran)后(hou)在(zai)第(di)(di)(di)(di)(di)(di)一、第(di)(di)(di)(di)(di)(di)二(er)孔(kong)(kong)中(zhong)加(jia)(jia)(jia)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)稀(xi)釋(shi)(shi)(shi)(shi)液(ye)50μl，混(hun)(hun)(hun)(hun)勻(yun)(yun)(yun)；然(ran)(ran)后(hou)從第(di)(di)(di)(di)(di)(di)一孔(kong)(kong)、第(di)(di)(di)(di)(di)(di)二(er)孔(kong)(kong)中(zhong)各(ge)取(qu)100μl分(fen)(fen)別(bie)加(jia)(jia)(jia)到(dao)第(di)(di)(di)(di)(di)(di)三(san)孔(kong)(kong)和第(di)(di)(di)(di)(di)(di)四(si)孔(kong)(kong)，再在(zai)第(di)(di)(di)(di)(di)(di)三(san)、第(di)(di)(di)(di)(di)(di)四(si)孔(kong)(kong)分(fen)(fen)別(bie)加(jia)(jia)(jia)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)稀(xi)釋(shi)(shi)(shi)(shi)液(ye)50μl，混(hun)(hun)(hun)(hun)勻(yun)(yun)(yun)；然(ran)(ran)后(hou)在(zai)第(di)(di)(di)(di)(di)(di)三(san)孔(kong)(kong)和第(di)(di)(di)(di)(di)(di)四(si)孔(kong)(kong)中(zhong)先(xian)各(ge)取(qu)50μl棄掉(diao)，再各(ge)取(qu)50μl分(fen)(fen)別(bie)加(jia)(jia)(jia)到(dao)第(di)(di)(di)(di)(di)(di)五(wu)、第(di)(di)(di)(di)(di)(di)六孔(kong)(kong)中(zhong)，再在(zai)第(di)(di)(di)(di)(di)(di)五(wu)、第(di)(di)(di)(di)(di)(di)六孔(kong)(kong)中(zhong)分(fen)(fen)別(bie)加(jia)(jia)(jia)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)稀(xi)釋(shi)(shi)(shi)(shi)液(ye)50ul，混(hun)(hun)(hun)(hun)勻(yun)(yun)(yun)；混(hun)(hun)(hun)(hun)勻(yun)(yun)(yun)后(hou)從第(di)(di)(di)(di)(di)(di)五(wu)、第(di)(di)(di)(di)(di)(di)六孔(kong)(kong)中(zhong)各(ge)取(qu)50μl分(fen)(fen)別(bie)加(jia)(jia)(jia)到(dao)第(di)(di)(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)(di)(di)八孔(kong)(kong)中(zhong)，再在(zai)第(di)(di)(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)(di)(di)八孔(kong)(kong)中(zhong)分(fen)(fen)別(bie)加(jia)(jia)(jia)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)稀(xi)釋(shi)(shi)(shi)(shi)液(ye)50μl，混(hun)(hun)(hun)(hun)勻(yun)(yun)(yun)后(hou)從第(di)(di)(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)(di)(di)八孔(kong)(kong)中(zhong)分(fen)(fen)別(bie)取(qu)50μl加(jia)(jia)(jia)到(dao)第(di)(di)(di)(di)(di)(di)九(jiu)、第(di)(di)(di)(di)(di)(di)十(shi)孔(kong)(kong)中(zhong)，再在(zai)第(di)(di)(di)(di)(di)(di)九(jiu)第(di)(di)(di)(di)(di)(di)十(shi)孔(kong)(kong)分(fen)(fen)別(bie)加(jia)(jia)(jia)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)稀(xi)釋(shi)(shi)(shi)(shi)液(ye)50μl，混(hun)(hun)(hun)(hun)勻(yun)(yun)(yun)后(hou)從第(di)(di)(di)(di)(di)(di)九(jiu)第(di)(di)(di)(di)(di)(di)十(shi)孔(kong)(kong)中(zhong)各(ge)取(qu)50μl棄掉(diao)。（稀(xi)釋(shi)(shi)(shi)(shi)后(hou)各(ge)孔(kong)(kong)加(jia)(jia)(jia)樣量都為50μl，濃度分(fen)(fen)別(bie)為1800ng/L，1200ng/L，600ng/L，300ng/L， 150ng/L）。

2. 加(jia)樣：分別設空(kong)(kong)白孔(kong)(kong)（空(kong)(kong)白對照孔(kong)(kong)不加(jia)樣品(pin)(pin)(pin)及(ji)酶(mei)標(biao)試劑，其余各步操作相(xiang)同）、待(dai)測樣品(pin)(pin)(pin)孔(kong)(kong)。在酶(mei)標(biao)包被板上(shang)待(dai)測樣品(pin)(pin)(pin)孔(kong)(kong)中先加(jia)樣品(pin)(pin)(pin)稀(xi)釋液40μl，然后再加(jia)待(dai)測樣品(pin)(pin)(pin)10μl（樣品(pin)(pin)(pin)最終稀(xi)釋度為5倍(bei)）。加(jia)樣將(jiang)樣品(pin)(pin)(pin)加(jia)于酶(mei)標(biao)板孔(kong)(kong)底部，盡量不觸及(ji)孔(kong)(kong)壁，輕輕晃動混(hun)勻。

3. 溫育：用封板(ban)膜封板(ban)后置37℃溫育30分鐘(zhong)。

4. 配(pei)液：將30（48T的(de)20倍）倍濃縮洗滌液用(yong)蒸餾水30（48T的(de)20倍）倍稀(xi)釋后(hou)備(bei)用(yong)。

5. 洗(xi)滌：小心揭掉封板膜，棄(qi)去液體，甩干，每(mei)孔加滿洗(xi)滌液，靜置(zhi)30秒后(hou)棄(qi)去，如此(ci)重復5次(ci)，拍干。

6. 加(jia)酶(mei)：每孔加(jia)入酶(mei)標試劑50μl，空白孔除外。

7. 溫(wen)育(yu)：用封(feng)板膜封(feng)板后置(zhi)37℃溫(wen)育(yu)30分鐘(zhong)。

8. 洗(xi)滌：小心揭(jie)掉封板膜，棄(qi)去液體，甩干，每孔加滿洗(xi)滌液，靜置30秒后棄(qi)去，如此重復5次，拍干。

9. 顯(xian)色：每(mei)孔先加入(ru)顯(xian)色劑A50μl，再加入(ru)顯(xian)色劑B50μl，輕輕震蕩(dang)混勻，37℃避(bi)光(guang)顯(xian)色15分鐘.

10. 終(zhong)止(zhi)：每孔加終(zhong)止(zhi)液(ye)50μl，終(zhong)止(zhi)反應（此時藍色立轉黃色）。

11. 測定(ding)：以(yi)空(kong)(kong)白空(kong)(kong)調(diao)零(ling)，450nm波長依(yi)序測量各孔的(de)吸(xi)光度(du)（OD值）。 測定(ding)應(ying)在加終止(zhi)液后15分鐘以(yi)內進行。

      以上是上海研(yan)域生物(wu)(wu)給大家介紹羊ELISA試(shi)劑(ji)盒的作(zuo)用，研(yan)域生物(wu)(wu)化學試(shi)劑(ji)有限(xian)公司宗旨是為了中國生命科學事業的騰飛，我們全力以赴，以更好的品質、更優惠(hui)的價格、更專(zhuan)業的技術(shu)服務好每一位客戶的需(xu)求。