Super ECL Detection Reagent ECL化学发光超敏显色试剂盒 [36208ES60]

Super ECL Detection Reagent ECL化学发光超敏显色试剂盒

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ECL化学发光超敏显色试剂盒用于检测直接或间接标记辣根过氧化物酶(HRP)的抗体及其关联的抗原。其原理是,蛋白质或核酸在电泳后转移到印迹膜上,以一抗及HRP标记的二抗结合膜上的目的蛋白,或以HRP标记的探针直接或间接结合膜上的核酸。洗膜后用本产品配制的ECL工作液,室温孵育膜数分钟,将印迹膜用保鲜膜包被粘贴固定于X光片曝光暗盒。然后转入暗室将X光胶片压在膜上曝光数秒到数小时,显影定影后蛋白质或核酸条带可清晰显示在X光胶片上。
本试剂盒采用了独特的发光底物系统,ECL化学发光超敏显色试剂盒是目前最灵敏的商业化荧光ECL检测试剂。
产品特点1.具有极高灵敏度和高信噪比,可检测低皮克级抗原;2.发光迅速,亮度强,可在日光灯下观察到印迹膜条带;3.持续发光时间长,荧光可使X光胶片感光达5小时以上,特别适用于痕量蛋白或核酸检测;4.可使用更高的抗体稀释倍数(12000~110000),大大节省抗体。
产品信息
组分编号 组分名称 36208ES60100 mL 36208ES76500 mL
36208-A A 50 mL 250 mL
36208-B B 50 mL 250 mL
 储存条件2~8避光保存。有效期1年
使用说明1. 执行常规电泳、转膜、HRP标记抗体或者HRP标记核酸探针孵育、洗膜。【注】:ECL发光液是HRP的显色底物,因此检测系统最终必须基于HRP酶标记抗体或者核酸探针。
2. 最后一次洗膜的同时,新鲜配制发光工作液:分别取等体积的A液和B液,混匀后,室温放置备用。【注】:建议立即使用工作液,室温放置数小时后仍可使用但灵敏度略有降低。
3. 用平头镊取出膜,搭在滤纸上沥干洗液,勿使膜完全干燥。将膜完全浸入发光工作液125μL发光工作液/cm2膜)中,与发光工作液充分接触。室温孵育3分钟,准备立即压片曝光。【注】:孵育时间过长不会增加灵敏度,有时还会导致曝光条带异常。发光过程的本质是酶促反应,使用过少的发光工作液不利反应进行,也会导致膜上条带曝光不均和明显降低灵敏度。为达节约目的可将膜剪小,但勿降低发光液使用比例。
4. 用镊子夹起膜,搭在滤纸上沥干发光工作液。但勿洗去发光液。
5. X光胶片暗盒内表面铺一张面积大于膜的保鲜膜。将印迹膜贴在保鲜膜上,将保鲜膜折起来完全包裹印迹膜,去除气泡和皱褶,可剪去边缘部多余的保鲜膜。用滤纸吸去多余的发光工作液。用胶带将覆盖印迹膜的保鲜膜固定在暗盒内,蛋白带面向上。
6. 暗房内压X光胶片,分别曝光不同的时间,如数秒到数分钟。显影冲洗。
注意事项1.步骤1~5可在日光灯下操作;但发光液曝露于强光下时间过久灵敏度可能略有降低,移到暗房操作可避免。戴手套可以避免在膜上留下手印,保持膜的干净。
2.长时间曝光或蛋白过量,将加深背景并使条带强弱变化失去线性关系。曝光不足则条带模糊。
3.发光工作液孵育约3分钟后膜上的条带发光。强条带发光在暗房中肉眼可见,低丰度蛋白条带发光较弱甚至肉眼不可见但可使X光胶片曝光。不能简单以肉眼观察判断条带发光时间。肉眼不可见的荧光实际上可持续数小时并使X光胶片感光,因此弱带可曝光1~10小时。如果曝光后条带不佳,可用洗膜缓冲液洗膜,重新孵育二抗,然后重新用ECL发光和曝光。
4.由于超敏发光液极其灵敏,强烈推荐大多数进口抗体起始浓度为一抗1∶1000~1∶4000,二抗1∶2000~1∶5000。抗体浓度过高将造成高背景或没有条带,导致失败。
5.某些保鲜膜包裹印迹膜时可能会淬灭荧光,应选择高质量保鲜膜。
6.使用肉眼可见的预染色蛋白Marker和荧光-放射自显影曝光标签可精确确定胶片上条带的位置和大小。
7.NaN3能抑制HRP活性,若回收HRP标记探针或者抗体应避免使用NaN3,如必需使用勿超过0.01%。
8.本品无特殊毒性,按普通化学品处理。
9.本产品仅作科研用途!
10.为了您的安全和健康,请穿实验服并佩戴一次性手套操作。
Ver.CN20230726

 
产品属性

保存条件:4ºC避光保存

保质期:有效期1年

产地:上海翌圣

英文名:Super ECL Detection Reagent

库存:大量

供应商:翌圣生物科技(上海)股份有限公司

CAS号:点击查看

货号:36208ES60

品牌:翌圣生物(Yeasen)

该产品被引用文献
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