Mab5718 sp

Total protein was prepared from liver tissues using radioimmunoprecipitation assay (RIPA) lysis buffer. Cell lysates were centrifuged at 12,000 rpm for 20 min, and protein concentrations were measured using the bicinchoninic acid (BCA) protein assay (DNAbiotech, Iran). Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride (PVDF) membrane. The membrane was blocked using 3% skim milk powder at 4°C overnight and then hybridized with the primary antibodies targeting the proteins, including GADPH and α-SMA (R&D, USA, catalog number MAB5718-SP and MA5-15871, respectively). For the appearance of the protein bands, they were incubated with secondary antibodies horseradish peroxidase (HRP) conjugate goat anti-mouse IgG1 (Invitrogen, USA, catalog number A10551) for 1 h at room temperature. The blotting bands were visualized using the ECL system (Merck, Germany).

Total protein was prepared from liver tissues using radioimmunoprecipitation assay (RIPA) lysis buffer. Cell lysates were centrifuged at 12,000 rpm for 20 min, and protein concentrations were measured using the bicinchoninic acid (BCA) protein assay (DNAbiotech, Iran). Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride (PVDF) membrane. The membrane was blocked using 3% skim milk powder at 4°C overnight and then hybridized with the primary antibodies targeting the proteins, including GADPH and α-SMA (R&D, USA, catalog number MAB5718-SP and MA5-15871, respectively). For the appearance of the protein bands, they were incubated with secondary antibodies horseradish peroxidase (HRP) conjugate goat anti-mouse IgG1 (Invitrogen, USA, catalog number A10551) for 1 h at room temperature. The blotting bands were visualized using the ECL system (Merck, Germany).