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Western blot有多种类型的膜可以用，不是所有类型的膜都适合做Stripping。 PVDF膜是最适合做Stripping的膜类型，蛋白样本的丢失量最小。 抗体清除液是什么?Western抗体清除液又称为一二抗清除液或膜再生液（stripping buffer），常用于WB中转移了蛋白的膜的重复利用。 对于曝光后效果较好的膜，先用TBST（Tris缓冲盐水+吐温）进行清洗，去除膜表面残留的试剂和杂质。 将清洗后的膜浸泡于甲醇中，浸泡时间控制在5秒左右，甲醇可快速固定膜上的蛋白，增强膜的稳定性。
a.加入适量蒸馏水装有NC膜或 PVDF膜 的抗体孵育盒中，漂洗5min（液体量能没过膜即可） b.弃掉蒸馏水加入适量抗体洗脱液，摇床上漂洗5min； 推荐先整膜转封闭完之后再裁膜，可以有效避免把胶切得稀碎，胶是脆的，膜是软的，所以建议切膜不切胶。 整膜封闭还有一个好处是比较节约封闭液。 A．用TBST一直洗膜，洗8hrs以上就可以清除多数蛋白的信号，信号过强的蛋白如内参除外。 所以，当你的抗体非常弱的时候，孵育过夜实际上相当于stripping，新的信号不会出现，原来的信号也会被漂洗干净，出现白板。
小编推荐使用配方三，优点是可以常温使用，并且适用于NC和PVDF膜。 根据小编经验，配方三比较温和，亲测一张膜可以反复剥离和敷抗体三次没问题。
由于最常用用于Western检测，而常被称作Western洗膜盒、抗体孵育盒等。 本产品为聚丙烯 (polypropylene)材质，与普通的1.5ml离心管的材质相同，确保与蛋白和核酸的非特异性吸附极低。 该方法通过洗脱已经孵育的抗体，可以将同一张膜用于重复检测多个靶蛋白，从而提高检测效率和节省成本。 以下是关于WB抗体洗脱重新孵育的详细介绍。 对于贴壁细胞：4℃ 预冷的 PBS 洗 2~3 遍，用细胞刮子刮下细胞，或用胰蛋白酶处理细胞使细胞不再贴壁很紧，并用移液器吹打下细胞。
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