产品简介 本产品采用温和洗涤配方，可在不影响目的蛋白的情况下，去除结合在印迹膜上的一抗和二抗，使同一张膜可进行多次抗体检测，无需反复电泳和转膜，节省样品和 时间，适用于使用NC或PVDF膜进行Western Blot检测时条件的优化或同一样品不同蛋 白的检测。 注意事项 1．建议先检测表达量较低的目的蛋白，用再生液处理后检测表达量高的蛋白，如内参 蛋白等。 2．请佩戴手套操作。 操作步骤 1．将曝光后的膜取出，加入适量的Stripping Buffer（蛋白印迹膜再生液），再生液充分覆盖膜表面，8.5 cm×5.5 cm膜加入15 mL左右蛋白印迹膜再生液，室温振摇孵育15分钟左右（孵育时间应根据不同的目的蛋白来调整：比如内参抗体等表达量较高的蛋白，使用蛋白印迹膜再生液时可以延长孵育时间至1小时或者在37℃孵育30分 钟）。 2．弃去蛋白印迹膜再生液，用15 mL缓冲液（PBST或TBST）洗膜3次，每次5 分钟， 室温振摇。 3．为了检测酶标二抗洗脱是否完全，此时可以用显色方法来确定二抗是否洗脱掉。 4．待检测完毕，确认膜上无残留的酶活性，再生后的膜通过加入15 mL的封闭液进行封闭，室温30分钟或者4℃封闭过夜。 5．重新加入待测一抗，进行下一轮的WB实验。 Products This product adopts a mild washing formula, which can remove the primary and secondary antibodies bound on the blotting membrane without affecting the target protein, so that the same membrane can be used for multiple antibody detection without repeated electrophoresis and membrane transfer, saving samples and time. It is suitable for optimizing the conditions of Western Blotting or detecting different proteins of the same sample using NC or PVDF membranes. Caveat 1. It is recommended to detect the target proteins with low expression first, and detect the proteins with high expression, such as the internal reference proteins, after treatment with regeneration solution. 2. Wear gloves when handling. Procedure 1. Remove the exposed membrane, add appropriate amount of Stripping Buffer (protein blotting membrane regeneration solution), the regeneration solution fully covers the surface of the membrane, 8.5 cm × 5.5 cm membrane add about 15 mL of the protein blotting membrane regeneration solution, incubate at room temperature with shaking for about 15 minutes (incubation time should be adjusted according to different target proteins: for example, internal reference antibody and other proteins with high expression, the incubation time can be extended to 1 hour or 30 minutes at 37°C when using the protein blotting membrane regeneration solution). (The incubation time should be adjusted according to different target proteins: for example, for proteins with higher expression such as endosome antibody, the incubation time can be extended to 1 hour or 30 minutes at 37℃ when using the protein blotting membrane regeneration solution). 2. Discard the protein blotting membrane regeneration solution, and wash the membrane with 15 mL of buffer (PBST or TBST) three times, each time for 5 minutes, shaking at room temperature. 3. In order to test whether the elution of the secondary antibody of the enzyme label is complete, the color development method can be used to determine whether the secondary antibody is eluted or not. 4. When the assay is completed, confirm that there is no residual enzyme activity on the membrane, and the regenerated membrane is closed by adding 15 mL of containment solution for 30 minutes at room temperature or overnight at 4℃. 5. Re-add the primary antibody to be tested and proceed to the next round of WB experiments.
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蛋白印迹膜再生液
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