| 货号 (SKU) | 包装规格 | 是否现货 | 价格 | 数量 |
|---|---|---|---|---|
| E266188-100ml |
100ml |
现货  |
| |
| E266188-500ml |
500ml |
现货 |
|
| 英文名称 | ECL Plus |
|---|---|
| 储存温度 | 2-8°C储存,避光,干燥 |
| 运输条件 | 冰袋运输 |
| 产品介绍 |
包装：100ml（50ml+50ml）,500ml（250ml+250ml） 产品描述 ECL Plus超敏发光液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。可通HRP实现低皮克级的免疫印迹检测。 （1）可使用更高的抗体稀释倍数(1:2000～1:10000)，极其节省抗体。 （2）简单易用— 可替代其它公司的ECL发光底物，操作步骤无需进行特别优化 （3）灵敏度更高— 可检测低皮克级的蛋白 （4）信号持续时间更长— 光信号持续时间长达5小时 （5）更多成像方法— 适用于X射线胶片、CCD或激光成像仪 （6）价格更经济 应用 用于HRP标记抗体的Western Blot和HRP标记探针的核酸杂交。 使用方法 1. 执行常规SDS-PAGE、转膜和Western Blot步骤。注意用HRP标记lgG或用一抗-链亲和素-生物素-HRP夹法。 操作概述 注：优化抗原和抗体的浓度。必须使用建议的抗体稀释度，以保证阳性结果。 1) 将一抗浓度稀释到0.05～1ug/ml 2) 将二抗浓度稀释到0.005～0.04ug/mL 3) 将两种底物组份按1:1比例混合，制备底物工作液。 注：暴漏于日光或任何其他强光可能损害工作液，为获得最佳结果，将此工作液保存在琥珀色瓶中，并避免长期暴漏于任何强光。短时间暴漏于实验室常规照明不会损害该工作液。 4) 将印迹膜在ECL底物工作液中孵育5分钟。 5) 吸出多余试剂。用清洁的塑料膜盖住该印迹膜。 6) 使印迹膜在X光胶片上曝光。 2. Western Blot最后一次洗膜的同时新鲜配制发光工作液：分别取等体积的溶液A和B，放入干净容器中混合。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. 用镊子取出膜，搭在滤纸上沥干洗液但勿使膜完全干燥。将膜完全浸入发光工作液（0.125mL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。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用量。 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. 打开X光胶片暗盒，在暗盒内表面铺一张面积大于膜的保鲜膜。将Western Blot膜贴在保鲜膜上，将保鲜膜折起来完全包裹Western Blot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Western Blot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。 储存 安全性 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之。戴手套可以避免在膜上留下手印。 （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，因而弱带可曝光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，回收第二抗体应避免使用NaN3，如必需使用勿超过0.01%。 ECL Plus Ultra Sensitive Luminescent Solution is used to detect antibodies that directly or indirectly label horseradish peroxidase HRP and its associated antigens. HRP can be used to achieve low picogram-level western blot detection. (1) A higher antibody dilution factor (1:2000～1:10000) can be used, which saves the antibody extremely. (2) Simple and easy to use-can replace other companies' ECL luminescent substrates, and the operation steps do not need to be specially optimized (3) Higher sensitivity-can detect low picogram protein (4) Longer signal duration-the light signal lasts up to 5 hours (5) More imaging methods-suitable for X-ray film, CCD or laser imager (6) The price is more economical Application The Western Blot for HRP-labeled antibodies and the nucleic acid hybridization of HRP-labeled probes. Instructions 1. Perform routine SDS-PAGE, transfer and Western Blot procedures. Note the use of HRP-labeled lgG or primary antibody-streptavidin-biotin-HRP clip method. Operation overview Note: Optimize the concentration of antigen and antibody. The recommended antibody dilution must be used to ensure a positive result. 1) Dilute the primary antibody concentration to 0.05～1ug/ml 2) Dilute the concentration of the secondary antibody to 0.005～0.04ug/mL 3) Mix the two substrate components at a ratio of 1:1 to prepare the substrate working solution. Note: Exposure to sunlight or any other strong light may damage the working fluid. For best results, store this working fluid in an amber bottle and avoid long-term exposure to any strong light. Exposure to routine lighting in the laboratory for a short time will not damage the working fluid. 4) Incubate the blot in the ECL substrate working solution for 5 minutes. 5) Aspirate the excess reagent. Cover the blotting membrane with a clean plastic film. 6) Expose the blot film on X-ray film. 2. Freshly prepare the luminescent working solution while washing the membrane for the last time by Western Blot: Take equal volumes of solutions A and B respectively, and mix them in a clean container. It is recommended to use the working solution immediately, it can still be used after a few hours at room temperature, but the sensitivity is slightly reduced. 3. Take out the membrane with tweezers, put it on the filter paper and drain the dry cleaning solution, but do not let the membrane dry completely. The film is completely immersed in the luminescent working fluid (0.125mL luminescent working fluid/cm2 film) and fully contacted with the luminescent working fluid. Incubate at room temperature for 3 minutes, ready to press and expose immediately. Too long incubation time will not increase sensitivity and sometimes cause abnormal exposure bands. The essence of the luminescence process is an enzymatic reaction. Using too little luminescence working fluid will adversely affect the progress of the reaction, and will also cause uneven exposure of the bands on the film and significantly reduce sensitivity. In order to achieve the purpose of saving, the film can be cut small but do not reduce the amount of luminescent liquid. 4. Use tweezers to pick up the film and place it on the filter paper to drain the luminous working solution. But do not wash off the luminous liquid. 5. Open the X-ray film cassette, and spread a fresh-keeping film with an area larger than the film on the inner surface of the cassette. Place the Western Blot film on the plastic wrap, fold the plastic wrap to completely wrap the Western Blot film, remove bubbles and wrinkles, and cut off the excess plastic wrap at the edges. Use filter paper to absorb excess luminescent working fluid. Fix the fresh-keeping film covering the Western Blot film in the cassette with tape, with the protein band facing up. 6. Press X-ray film in the darkroom and expose for different time, such as several seconds to several minutes. Develop and rinse. , Store Store at 4℃ sealed and protected from light for more than one year. Can be placed at room temperature for a short time. Safety No special toxicity, treat as common chemicals. Precautions (1) Steps 1 to 5 can be operated under a fluorescent lamp; however, the sensitivity may be slightly reduced when the luminescent liquid is exposed to strong light for too long, and it can be avoided by moving to a darkroom. Wearing gloves can avoid leaving fingerprints on the membrane. (2) Long-term exposure or excessive protein will deepen the background and make the band strength change lose the linear relationship. Underexposure will blur the bands. (3) The strips on the membrane glow after incubating for about 3 minutes in the luminescent working solution. The strong band luminescence is visible to the naked eye in the darkroom, and the low-abundance protein band luminescence is weak or even invisible to the naked eye, but it can be exposed to X-ray film. The light emission time of the strip cannot be judged simply by visual observation. Fluorescence that is invisible to the naked eye can actually last for several hours and sensitize the X-ray film, so weak bands can be exposed for 1-10 hours. If the band is not good after exposure, wash the membrane with membrane washing buffer, re-incubate the secondary antibody, and then re-illuminate and expose with ECL. (4) As the ultra-sensitive luminescent solution is extremely sensitive, it is strongly recommended that the initial concentration of most imported antibodies is 1:1000～1:4000 for the primary antibody and 1:2000～1:5000 for the secondary antibody. Too high antibody concentration will cause high background or no bands, leading to failure. (5) Some plastic wrap may quench the fluorescence when wrapping the blotting membrane, so high-quality plastic wrap should be selected. (6) Pre-stained protein Marker and fluorescence-autoradiography exposure label can be used to accurately determine the position and size of the band on the film. (7) NaN3 can inhibit the activity of HRP. The use of NaN3 should be avoided when recovering the secondary antibody. If necessary, use no more than 0.01%. |
| 敏感性 | 对光线敏感 |
|---|
¥4,562.90
¥1,667.90
¥1,999.90
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