Xiang, Yufang, et al. Scientific reports, 2021, 11(1), 12022.
The choice of blotting membrane is one of the key factors affecting the sensitivity of protein detection in the transfer from gel to membrane in western blotting (WB). This work compares the performance of polyvinylidene fluoride (PVDF) and nitrocellulose (NC) membranes for WB.
Research methods and results
· The binding capacity of these two membranes for proteins or glycoproteins in relation to the molecular weight of the target protein was investigated by incubating the proteins transferred to PVDF or NC membranes with a series of antibodies and different types of lectins, and the antibody re-probing capacity of these two membranes was also explored.
· In addition, the characterization of protein binding to PVDF or NC membranes was described from the results of directly incubating a series of proteins of different molecular weights with PVDF or NC membranes, and the transfer time for different antigens was investigated.
· The results showed that NC membrane had a better binding capacity for small molecular weight proteins than PVDF membrane, while PVDF membrane had a greater advantage in binding capacity for high molecular weight proteins and glycoproteins, while there was no difference between the two membranes in terms of binding capacity for medium molecular weight proteins. Moreover, considering the corresponding transfer time, when the molecular weight of the protein is small, the transfer time can be appropriately shortened, otherwise it can be appropriately extended to allow more proteins to be retained on the electroblotting membrane.
