Create a step-by-step Western Blot protocol figure: (1) protein extraction with RIPA buffer + protease/phosphatase inhibitors, (2) BCA protein quantification with standard curve, (3) SDS-PAGE sample preparation (reducing, boiling 95°C 5min), (4) gel electrophoresis (stacking 4%, resolving 10% polyacrylamide), (5) semi-dry transfer to PVDF membrane (25V, 30min), (6) Ponceau S total protein staining verification, (7) blocking (5% BSA/TBST, 1h RT), (8) primary antibody incubation (4°C overnight), (9) HRP-conjugated secondary antibody (1h RT), (10) ECL detection and digital imaging. Show expected band patterns.
