How to distinguish non-specific bands from my bands in Western blot?

1． There is a relatively large possibility that this is your band, protein may be methylated, acetylated, phosphorylated and other modifications after expression to make its molecular weight become larger, you can consult the relevant literature, if the size of the protein in the literature is consistent with your protein size, the band is your destination band.  

2． You can go to the antibody instructions or the official website to view the non-specific bands caused by the antibody used, combined with some of the non-specific bands you find when you do experiments, it is basically the non-specific bands caused by the antibody.  

3． Check whether the marker is standard, you can add some proteins that determine the molecular weight when doing experiments to see if the results are correct to determine whether the marker is standard.

4． If there is more negative charge on the protein, its migration speed will be slower.