Description
ProductsThis 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. Caveat1. 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. Procedure1. 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