Western Blot Protein Detection Protocol

Via SDS acrylamide gel electrophoresis, protein sample was transferred to nitrocellulose filter membrane (10600002, Whatman) and then blocked in 5% skimmed milk for 1 h. The membrane was then incubated with primary antibodies overnight at 4°C and then incubated with secondary antibody for 1 h. Antibodies used in this study are listed in Table 1. Odyssey Infrared Imaging System (LI-COR Biosciences, Lincoln, NE, United States) and ECL Imaging System (610007-8Q, Clinx Science Instruments Co., Ltd.) were used for visualization of protein bands. Quantitative analysis of blots was performed using ImageJ software (Fiji) (Li et al., 2021 (link)).