The eluent of primary and secondary antibodies, also known as primary and secondary antibody removal solution or membrane regeneration solution, can be used for multiple Western Blot tests after elution of the membrane with transferred proteins in the Western experiment. For example, after primary and secondary antibody binding and subsequent chemiluminescence detection in Western, GAPDH, β-actin and other internal reference proteins can be detected by primary and secondary antibody eluent treatment. For example, the number of protein samples extracted is too small for multiple protein detection, and the antibody binding on the membrane can also be removed by primary and secondary antibody eluent treatment, and multiple detection can be carried out. This strategy can eliminate costly steps such as protein electrophoresis and membrane transfer, and can eliminate errors caused by re-loading, so that the results are more comparable.
The main ingredients of this product are Tris-HCl buffer, SDS and β-mercaptoethanol, which are acidic.
Primary and secondary antibody eluent
1. The membrane to be treated was rinsed with distilled water for 5min, and then fully immersed in the eluent of primary and secondary reagents. The membrane was shaken on a shaker at room temperature for 10-60 min. After eluting for a certain time, ECL color and exposure detection can be carried out to determine whether the membrane is eluted clean. If the elution is not clean, then continue to enter the eluent of primary and secondary antibodies to soak until elution is clean.
2. After elution, the membrane was removed and rinsed twice with TBST, and the primary antibody was directly incubated for detection.
1. This reagent is suitable for films detected by chemiluminescent reagents such as ECL. Western detection by non-chemiluminescent reagents such as DAB, NBT/BCIP, etc., is not suitable for this product.
2. Membrane type, antibody and antigen characteristics may affect the elution degree, so it is recommended to conduct fumbling experiment to obtain the best conditions.
3. The film sealed with skim milk powder is easier to regenerate than the film sealed with BSA, so skim milk powder G5002 is recommended for sealing.
4. It is recommended to use PVDF membranes for best results.
5. This product is prone to white flocculent precipitation at low temperature, which is a normal phenomenon. It can be used after reheating at 37℃.
6. Please wear lab clothes and disposable gloves during operation.