ECL chemiluminescence is currently the most commonly used and sensitive Western Blot assay. Since these substrates do not precipitate on the membrane surface and are firmly bonded to the membrane, the affinity-bound primary and secondary antibodies can be removed by a first anti-secondary stripping buffer. The primary antibody secondary antibody removal solution is sufficiently strong to effectively dissociate the bound antibody, and is also mild enough to allow the transferred target protein to remain intact on the NC membrane or PVDF membrane. By using the primary anti-secondary antibody removal solution and fully removing the primary antibody, it is very convenient to reuse the used membrane to detect other proteins. Compared with re-running SDS-PAGE glue, it not only saves time and effort, but also eliminates the error caused by re-loading, making comparability stronger.

First, the method of use

1. Cover the ECL-emitting membrane with this product and incubate for 5 - 10 minutes at room temperature.

2. Discard the removal solution, then add fresh removal solution to cover the ECL-emitting membrane and incubate for 5-10 minutes at room temperature.

3. Discard the removal solution and wash twice with PBS for 10 minutes.

4. Wash TBST for 5 minutes. Can be used for the next closure, incubation.

Second, matters needing attention

1. If horseradish peroxidase (HRP) is used, skim milk powder or BSA should be used when blocking. If alkaline phosphatase is used, casein should be used when blocking.

2. For best results, PVDF membranes are recommended. However, the nitrocellulose membrane can also use the Western primary antibody secondary antibody removal solution.

3. This product is suitable for Western Blot detection using ECL or similar chemiluminescent reagents. Western Blot detection using non-chemiluminescent reagents, such as DAB, NBT/BCIP, etc., is not applicable to this product.

4. This Western primary antibody secondary antibody removal solution is slightly corrosive, please pay attention to protection when using.

5. It is difficult to say which Western Blot primary antibody secondary antibody removal solution works best for specific antigens and antibodies. In the case of poor primary antibody secondary removal, it may be considered to appropriately extend the incubation time of the primary antibody secondary antibody removal solution while ensuring that the incubation temperature is not lower than 20 °C. When the temperature is too low, the removal effect of the primary antibody will decrease. At the same time, you can also consider other primary anti-secondary solution removal solution to find the best Western Blot primary antibody secondary antibody removal solution suitable for your experimental conditions.

6. For your safety and health, wear protective clothing such as protective clothing, gloves, and masks.

Third, product information

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