1. Sample Preparation: Tissue samples are lysed to extract proteins, which are then quantified. 2. Gel Electrophoresis: Proteins are separated based on size using SDS-PAGE. 3. Transfer to Membrane: Proteins are transferred to a nitrocellulose or polyvinylidene fluoride (PVDF) membrane. 4. Blocking: The membrane is blocked to prevent non-specific binding. 5. Primary Antibody Incubation: The membrane is incubated with a primary antibody specific to the target protein. 6. Secondary Antibody Incubation: A secondary antibody, conjugated to a detection enzyme or fluorophore, is applied. 7. Detection: The target protein is visualized using chemiluminescent, fluorescent, or colorimetric methods.
