Western Primary&Secondary Antibody Removal Solution(Strong Alkaline)

Membrane regeneration fluids, also referred to as Western primary and secondary Stripping fluids or Western Stripping buffer, play a crucial role in the recycling of membranes used for protein transfer in Western blotting. In Western blotting, after the primary and secondary antibodies have bound to the target proteins and chemiluminescence detection has taken place, it becomes necessary to detect certain reference proteins with stable expression levels, such as GAPDH and β-actin, which serve as internal controls. The stripping buffer effectively removes the bound primary and secondary antibodies from the membrane, allowing the membrane to be reused for the detection of other proteins. However, it should be noted that reusing the same membrane multiple times may result in decreased protein signaling, thereby affecting the accuracy of the results. Therefore, it is generally recommended to reuse the membrane for two to three rounds before considering replacement. It is important to avoid overusing the stripping buffer as it may not be suitable for repeated reuse.
By utilizing primary and secondary antibody removers to completely strip off primary and secondary antibodies, it is feasible to conveniently reuse the utilized membrane for the detection of different proteins. This practice not only saves valuable time and effort but also circumvents any potential errors that may arise from reloading, thus enhancing the overall comparability. In contrast to rerunning a SDS-PAGE gel, the removal technique presents a more efficient and reliable alternative.
The Western primary and secondary antibodies remover is a strongly alkaline solution that works by breaking the bond between antibodies and the membrane. This process is done without compromising the protein transferred onto the membrane and is ideal for protein membrane reuse detected by Western chemiluminescence, especially PVDF and nitrate cellulose membranes. It is not recommended to use this reagent for Western tests with non-chemiluminescent reagents, like DAB, NBT/BCIP.

Notes:
If you are using horseradish peroxidase labeled antibodies in your experiment, it is important to prepare the blocking solution with 5% skimmed milk powder. On the other hand, for experiments using alkaline phosphatase labeled antibodies, the blocking solution should be prepared using casein. Proper preparation of the blocking solution is crucial for ensuring optimal performance of the labeled antibodies. By following these guidelines, you can expect reliable and accurate results from your experiments.
The alkaline-based Western primary and secondary antibody remover (2) is specifically designed for the efficient regeneration of PVDF membranes. It exhibits superior performance in removing antibodies from PVDF membranes compared to nitrate cellulose membranes.
The use of Western primary and secondary anti-eluting solution (strong alkaline) is ideal for primary and secondary anti-elution post detection of ECL and other similar chemiluminescent liquids. However, reagents that are not chemiluminescent such as DAB and NBT/BCIP cannot be used with this particular reagent. It is important to bear this in mind when conducting Western tests.
4, Western primary and secondary resistance remover (strong alkaline) is slightly corrosive, please pay attention to protection when using.
5. For your safety and health, please wear lab clothes and disposable gloves for operation.