How to perform western blot of a small protein?

1. The amount of loading, usually Western Blot per well loading 30-50μg protein, when the molecular weight is small, it is recommended to appropriately increase the amount of loading  

2. Electrophoresis, according to the protein molecular weight, the smaller the molecular weight, the larger the gel concentration should be selected, and pay attention to the process of running the gel when performing gel electrophoresis, so as not to run out of the gel with the protein, and you can stop running the gel when bromophenol blue runs out of the gel.

3. Transfer, it is recommended to use 0.2 μm PVDF membrane when transferring, before transferring the membrane, soak the PVDF membrane and gel with a transfer buffer to improve the transfer efficiency. The transfer time of small molecule protein can be appropriately reduced, about 40-50 minutes, and the transfer effect can be determined by Lichun red staining after the transfer.  

4. Antibodies, when using small molecule proteins for experiments, a lower antibody dilution concentration should be selected, which will help the binding of antibodies to proteins.  

 Hope these can help you!