雙氯青霉素檢測試劑盒規(guī)    格：96T

檢測標(biāo)本：各種動物肌肉組織、肝臟、尿液、血清、水產(chǎn)、蜜蜂、牛奶、糧食作物（玉米、大米、豆類、花生、麥類等）、飼料等

檢測方法：ELISA

檢測類型：酶聯(lián)免疫分析法

產(chǎn)品的用途：僅供科研究課題使用 

雙氯青霉素檢測試劑盒價格及詳細(xì)資料：電議,或咨詢在線客服,或者以郵件形式發(fā)到我司qysw@qiyibio.com .齊一生物科技（上海）有限公司提供的ELISA試劑盒受到了廣大科研單位的*肯定和認(rèn)同。*保證，價格公道，傾力為國內(nèi)外科研院校實驗室提供的產(chǎn)品。若有需要，我司將竭誠為您服務(wù)！

                 本試劑盒用于測定各種動物肌肉組織、肝臟、尿液、血清、飼料等水產(chǎn)/蜂蜜/牛奶相關(guān)液體樣本中殘留的定量檢測。 

實驗原理：

   本試劑盒應(yīng)用雙抗體夾心法測定標(biāo)本中該產(chǎn)品水平。用純化的本產(chǎn)品抗體包被微孔板，制成固相抗體，往包被單抗的微孔中依次加入本產(chǎn)品抗原，再與HRP標(biāo)記的本產(chǎn)品抗體結(jié)合，形成抗體-抗原-酶標(biāo)抗體復(fù)合物，經(jīng)過*洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍(lán)色，并在酸的作用下轉(zhuǎn)化成zui終的黃色。顏色的深淺和樣品中的本產(chǎn)品呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度（OD值），通過標(biāo)準(zhǔn)曲線計算樣品中該產(chǎn)品濃度。

試劑盒組成：

標(biāo)本要求： 

1．標(biāo)本采集后盡早進(jìn)行提取，提取按相關(guān)文獻(xiàn)進(jìn)行，提取后應(yīng)盡快進(jìn)行實驗。若不能馬上進(jìn)行試驗，可將標(biāo)本放于-20℃保存，但應(yīng)避免反復(fù)凍融

2．不能檢測含NaN3的樣品，因NaN3抑制辣根過氧化物酶的（HRP）活性。

操作步驟：

• 標(biāo)準(zhǔn)品的稀釋與加樣：在酶標(biāo)包被板上設(shè)標(biāo)準(zhǔn)品孔10孔，在*、第二孔中分別加標(biāo)準(zhǔn)品100μl，然后在*、第二孔中加標(biāo)準(zhǔn)品稀釋液50μl，混勻；然后從*孔、第二孔中各取100μl分別加到第三孔和第四孔，再在第三、第四孔分別加標(biāo)準(zhǔn)品稀釋液50μl，混勻；然后在第三孔和第四孔中先各取50μl棄掉，再各取50μl分別加到第五、第六孔中，再在第五、第六孔中分別加標(biāo)準(zhǔn)品稀釋液50ul，混勻；混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中，再在第七、第八孔中分別加標(biāo)準(zhǔn)品稀釋液50μl，混勻后從第七、第八孔中分別取50μl加到第九、第十孔中，再在第九第十孔分別加標(biāo)準(zhǔn)品稀釋液50μl，混勻后從第九第十孔中各取50μl棄掉。（稀釋后各孔加樣量都為50μl，濃度分別為1200 ng/L，800 ng/L ，400 ng/L，200ng/L， 100 ng/L）。
• 加樣：分別設(shè)空白孔（空白對照孔不加樣品及酶標(biāo)試劑，其余各步操作相同）、待測樣品孔。在酶標(biāo)包被板上待測樣品孔中先加樣品稀釋液40μl，然后再加待測樣品10μl（樣品zui終稀釋度為5倍）。加樣將樣品加于酶標(biāo)板孔底部，盡量不觸及孔壁，輕輕晃動混勻。
• 溫育：用封板膜封板后置37℃溫育30分鐘。
• 配液：將30（48T的20倍）倍濃縮洗滌液用蒸餾水30（48T的20倍）倍稀釋后備用。
• 洗滌：小心揭掉封板膜，棄去液體，甩干，每孔加滿洗滌液，靜置30秒后棄去，如此重復(fù)5次，拍干。
• 加酶：每孔加入酶標(biāo)試劑50μl，空白孔除外。
• 溫育：操作同3。
• 洗滌：操作同5。
• 顯色：每孔先加入顯色劑A50μl，再加入顯色劑B50μl，輕輕震蕩混勻，37℃避光顯色15分鐘. 
• 終止：每孔加終止液50μl，終止反應(yīng)（此時藍(lán)色立轉(zhuǎn)黃色）。
• 測定：以空白空調(diào)零，450nm波長依序測量各孔的吸光度（OD值）。 測定應(yīng)在加終止液后15分鐘以內(nèi)進(jìn)行。

注意事項：

• 試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用，酶標(biāo)包被板開封后如未用完，板條應(yīng)裝入密封袋中保存。
• 濃洗滌液可能會有結(jié)晶析出，稀釋時可在水浴中加溫助溶，洗滌時不影響結(jié)果。
• 各步加樣均應(yīng)使用加樣器，并經(jīng)常校對其準(zhǔn)確性，以避免試驗誤差。一次加樣時間控制在5分鐘內(nèi)，如標(biāo)本數(shù)量多，使用排槍加樣。
• 請每次測定的同時做標(biāo)準(zhǔn)曲線，做復(fù)孔。如標(biāo)本中待測物質(zhì)含量過高（樣本OD值大于標(biāo)準(zhǔn)品孔*孔的OD值），請先用樣品稀釋液稀釋一定倍數(shù)（n倍）后再測定，計算時請zui后乘以總稀釋倍數(shù)（×n×5）。
• 封板膜只限一次性使用，以避免交叉污染。
• 底物請避光保存。
• 嚴(yán)格按照說明書的操作進(jìn)行，試驗結(jié)果判定必須以酶標(biāo)儀讀數(shù)為準(zhǔn).
• 所有樣品，洗滌液和各種廢棄物都應(yīng)按傳染物處理。
• 本試劑不同批號組分不得混用。

10. 如與英文說明書有異，以英文說明書為準(zhǔn)。

計算：

以標(biāo)準(zhǔn)物的濃度為橫坐標(biāo)，OD值為縱坐標(biāo)，    

在坐標(biāo)紙上繪出標(biāo)準(zhǔn)曲線，根據(jù)樣品的OD      

值由標(biāo)準(zhǔn)曲線查出相應(yīng)的濃度；再乘以稀釋      

倍數(shù)；或用標(biāo)準(zhǔn)物的濃度與OD值計算出標(biāo)      

準(zhǔn)曲線的直線回歸方程式，將樣品的OD值      

代入方程式，計算出樣品濃度，再乘以稀釋

倍數(shù)，即為樣品的實際濃度。

試劑盒性能：

1.樣品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.95以上。

2.批內(nèi)與批見應(yīng)分別小于9%和11%     

保存條件及有效期：

1.試劑盒保存；2-8℃。

2．有效期：6個月

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