操作步驟：

1. 標(biao)準(zhun)品(pin)的稀(xi)(xi)釋(shi)與加(jia)樣(yang)：在酶標(biao)包被板(ban)上設(she)標(biao)準(zhun)品(pin)孔(kong)(kong)(kong)(kong)10孔(kong)(kong)(kong)(kong)，在di一、第(di)(di)(di)(di)二(er)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)分(fen)別加(jia)標(biao)準(zhun)品(pin)100μl，然(ran)后(hou)在di一、第(di)(di)(di)(di)二(er)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)加(jia)標(biao)準(zhun)品(pin)稀(xi)(xi)釋(shi)液(ye)50μl，混(hun)勻(yun)；然(ran)后(hou)從di一孔(kong)(kong)(kong)(kong)、第(di)(di)(di)(di)二(er)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)各(ge)取(qu)(qu)(qu)(qu)100μl分(fen)別加(jia)到第(di)(di)(di)(di)三孔(kong)(kong)(kong)(kong)和第(di)(di)(di)(di)四孔(kong)(kong)(kong)(kong)，再(zai)在第(di)(di)(di)(di)三、第(di)(di)(di)(di)四孔(kong)(kong)(kong)(kong)分(fen)別加(jia)標(biao)準(zhun)品(pin)稀(xi)(xi)釋(shi)液(ye)50μl，混(hun)勻(yun)；然(ran)后(hou)在第(di)(di)(di)(di)三孔(kong)(kong)(kong)(kong)和第(di)(di)(di)(di)四孔(kong)(kong)(kong)(kong)中(zhong)(zhong)先各(ge)取(qu)(qu)(qu)(qu)50μl棄掉，再(zai)各(ge)取(qu)(qu)(qu)(qu)50μl分(fen)別取(qu)(qu)(qu)(qu)50μl分(fen)別加(jia)到第(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)八(ba)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)，再(zai)在第(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)八(ba)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)分(fen)別加(jia)標(biao)準(zhun)品(pin)稀(xi)(xi)釋(shi)液(ye)50μl，混(hun)勻(yun)后(hou)從第(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)八(ba)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)加(jia)到第(di)(di)(di)(di)五(wu)、第(di)(di)(di)(di)六(liu)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)，再(zai)在第(di)(di)(di)(di)五(wu)、第(di)(di)(di)(di)六(liu)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)分(fen)別加(jia)標(biao)準(zhun)品(pin)稀(xi)(xi)釋(shi)液(ye)50ul，混(hun)勻(yun)；混(hun)勻(yun)后(hou)從第(di)(di)(di)(di)五(wu)、第(di)(di)(di)(di)六(liu)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)各(ge)分(fen)別取(qu)(qu)(qu)(qu)50μl加(jia)到第(di)(di)(di)(di)九、第(di)(di)(di)(di)十(shi)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)，再(zai)在第(di)(di)(di)(di)九第(di)(di)(di)(di)十(shi)孔(kong)(kong)(kong)(kong)分(fen)別加(jia)標(biao)準(zhun)品(pin)稀(xi)(xi)釋(shi)液(ye)50μl，混(hun)勻(yun)后(hou)從第(di)(di)(di)(di)九第(di)(di)(di)(di)十(shi)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)各(ge)取(qu)(qu)(qu)(qu)50μl棄掉。（稀(xi)(xi)釋(shi)后(hou)各(ge)孔(kong)(kong)(kong)(kong)加(jia)樣(yang)量都(dou)為50μl，濃度分(fen)別為180 ng/L，120 ng/L ，60 ng/L，30 ng/，15 ng/L）。

2. 加樣：分(fen)別設空白(bai)孔（空白(bai)對照(zhao)孔不加樣品(pin)(pin)(pin)及酶(mei)標試劑，其余各步操作相同）、待(dai)測樣品(pin)(pin)(pin)孔。在酶(mei)標包被板上待(dai)測樣品(pin)(pin)(pin)孔中先加樣品(pin)(pin)(pin)稀(xi)釋(shi)液40μl，然后再(zai)加待(dai)測樣品(pin)(pin)(pin)10μl（樣品(pin)(pin)(pin)最終稀(xi)釋(shi)度為5倍）。加樣將樣品(pin)(pin)(pin)加于酶(mei)標板孔底部(bu)，盡量不觸及孔壁，輕輕晃動混勻(yun)。

3. 溫育(yu)：用封(feng)板(ban)(ban)膜封(feng)板(ban)(ban)后置(zhi)37℃溫育(yu)30分鐘(zhong)。

4. 配液(ye)：將30（48T的(de)20倍(bei)）倍(bei)濃(nong)縮(suo)洗滌液(ye)用蒸餾水30（48T的(de)20倍(bei)）倍(bei)稀釋(shi)后備用。

5. 洗滌(di)(di)：小心揭(jie)掉封(feng)板膜(mo)，棄(qi)去液體，甩干(gan)(gan)，每孔加滿洗滌(di)(di)液，靜(jing)置30秒后棄(qi)去，如此重復5次，拍干(gan)(gan)。

6. 加酶：每孔(kong)加入酶標試劑50μl，空白孔(kong)除(chu)外。

7. 溫育(yu)：操(cao)作(zuo)同(tong)3。

8. 洗滌(di)：操作同5。

9. 顯(xian)色(se)(se)：每孔先加入顯(xian)色(se)(se)劑A50μl，再加入顯(xian)色(se)(se)劑B50μl，輕輕震蕩混(hun)勻(yun)，37℃避光顯(xian)色(se)(se)15分(fen)鐘.

10. 終(zhong)止(zhi)(zhi)：每孔加終(zhong)止(zhi)(zhi)液(ye)50μl，終(zhong)止(zhi)(zhi)反應（此時藍(lan)色立轉(zhuan)黃(huang)色）。

11. 測(ce)定(ding)：以空白(bai)空調零，450nm波長依序測(ce)量各孔的(de)吸光度（OD值）。 測(ce)定(ding)應在加終止液后15分鐘(zhong)。雌二醇單克隆抗體

雌三醇單克隆抗體

HRP標(biao)記(ji)雌三醇單克隆抗(kang)體

甲基睪酮單克隆抗體

喹乙醇單克隆抗體

三聚氰胺單克隆抗體

賽庚啶多克隆抗體

金剛烷胺多克隆抗體

B1檢測試劑盒

M1檢測試劑盒

玉米赤(chi)霉烯(xi)酮(tong)檢測試劑盒

T-2毒素單克隆抗(kang)體

嘔吐毒素單克隆抗體

鹽酸克倫特羅檢測試劑盒

萊克多巴(ba)胺檢測試劑盒