普美克生產(chǎn)的超敏ECL發(fā)光底物檢測試劑盒用于檢測直接或間接標記辣根過氧化物酶（HRP）的抗體及其關聯(lián)的抗原。其原理是，蛋白質或核酸在電泳后轉移到印跡膜上，以一抗及HRP標記的二抗結合膜上的目的蛋白，或以HRP標記的探針直接或間接結合膜上的核酸。洗膜后用本產(chǎn)品配制的ECL工作液，室溫孵育膜數(shù)分鐘，將印跡膜用保鮮膜包被粘貼固定于X光片曝光暗盒。然后轉入暗室將X光膠片壓在膜上曝光數(shù)秒到數(shù)小時，顯影定影后蛋白質或核酸條帶可清晰顯示在X光膠片上。
本產(chǎn)品采用獨特的發(fā)光底物系統(tǒng)，降低曝光背景的同時引入新型的氧化劑，大大提高試劑盒的穩(wěn)定性，使其在室溫能穩(wěn)定放置一年。本產(chǎn)品配制的ECL工作液衰減較慢，15min內(nèi)熒光幾無減弱，檢測時產(chǎn)生的非特異背景非常低，也可節(jié)省抗體和待測樣品的用量，該試劑可用于PVDF膜和NC膜(硝酸纖維素膜)的Western blot免疫印跡等印跡發(fā)光顯色。除用于X光膠片，還可直接使用熒光CCD掃描。
1、為了您的安全和健康，請穿實驗服并戴一次性手套操作。
2、本產(chǎn)品A液和B液均對人體有害，操作時請小心，并注意有效防護避免直接接觸人體或吸入體內(nèi)。本產(chǎn)品僅限于專業(yè)人員的科學研究用，不得用于臨床診斷或治療，不得用于食品或藥品，不得存放于普通住宅內(nèi)。
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答：可能的原因有：
1、目的蛋白有多個修飾位點，本身可以呈現(xiàn)多條帶，建議查閱文獻或進行生物信息學分析，獲得蛋白序列的修飾位點信息，通過去修飾確定蛋白實際大小；
2、樣本處理過程中目的蛋白發(fā)生降解,建議加入蛋白酶抑制劑；樣本處理時在冰上操作；
3、雜蛋白多，建議處理目的蛋白；
4、抗體特異性不強，建議使用特異性強的抗體；
5、抗體孵育時間過久，建議減少抗體孵育時間；
6、二抗與抗原有交叉反應，建議選擇合適的封閉物；
7、二聚體或多聚體存在，建議增加蛋白質變性過程及強度；
8、底物顯色與曝光時間過長，建議縮短顯色及曝光的時間。
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