Absin ECL化學發(fā)光檢測試劑盒產品詳情
1. 基本屬性
中文名稱 | ECL化學發(fā)光檢測試劑盒 | ||
英文名字 | ECL luminescence reagent | ||
別名 | ECL發(fā)光液;HRP化學發(fā)光底物 | ||
產品貨號 | abs920 | ||
產品規(guī)格 | 2*25ml,2*50ml,2*250ml | ||
產品組份 | A液;B液 | ||
保存建議 | 2-8℃避光保存6個月。 | ||
產品說明書 |
2. 產品描述
Western blot底物發(fā)光檢測試劑可由標記于二抗上的辣根過氧化酶催化,產生化學發(fā)光反應,可以靈敏地檢測出目的蛋白的存在。ECL底物化學發(fā)光檢測試劑盒基于新一代增強型化學發(fā)光底物研制而成,并對成份做了優(yōu)化。產品背景低,穩(wěn)定性好。它由辣根過氧化物酶(HRP)催化發(fā)生化學反應,發(fā)出熒光,可對X光膠片曝光,也可直接進行l(wèi)uminometer檢測或者熒光CCD掃描。
3. 操作步驟
1. 按常規(guī)Western blot操作,二抗孵育后,進行最后一次洗滌時,根據(jù)膜的大小,按每10 cm2 膜混合0.5ml溶液A和0.5ml溶液B混勻,配制成發(fā)光檢測工作液。
2. 用平頭鑷子將膜取出,膜的下緣輕輕接觸吸水紙,以去除膜上多余的液體。膜的蛋白面朝上,置于潔凈保鮮膜(某些市售保鮮膜包裹印跡膜時可能會淬滅熒光,應選擇高質量保鮮膜)上。用吸管將配制的發(fā)光檢測液轉移到蛋白膜上,使其均勻覆蓋,室溫孵育2分鐘。
3. 用平頭鑷夾持蛋白膜,膜的下緣輕輕接觸吸水紙,以去除膜上多余的液體。膜的蛋白面朝上,包裹于潔凈保鮮膜內。輕輕趕出其間的氣泡,固定在X片暗盒內。
4. 在暗室中取一張X片置于包裹的膜上,合上暗盒,曝光30秒至1分鐘。立即顯影定影,根據(jù)其曝光強度,縮短或延長下一張X片的曝光時間(對微弱信號,曝光時間可延長至數(shù)小時)。
4. 用法說明
1.ECL A液和B液在吸取過程中必須要更換槍頭,A液和B液相互污染后會導致 A液或B液逐漸失效,影響后續(xù)的使用效果。
2.各溶液使用后,請蓋緊瓶蓋,以防失效。特別是B液,含有氧化劑,比較容易 被還原而失效。
3.ECL的熒光持續(xù)時間很長,但開始反應后的30分鐘內熒光更強一些,隨后熒光會逐漸減弱,因此請注意充分利用這熒光較強的30分鐘進行壓片。
4.ECL A液和B液均對人體有害,操作時請注意適當防護。 為了您的安全和健康,請穿實驗服并戴一次性手套操作。
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