The process of western blotting involves several steps: 1. Protein Extraction: Proteins are extracted from cells or tissues. 2. Gel Electrophoresis: Extracted proteins are separated based on size using SDS-PAGE. 3. Transfer: Separated proteins are transferred onto a membrane (usually PVDF or nitrocellulose). 4. Blocking: The membrane is blocked to prevent non-specific binding. 5. Antibody Incubation: The membrane is incubated with a primary antibody specific to the target protein, followed by a secondary antibody that is linked to a detection method. 6. Detection: Visualization of the target protein through chemiluminescence, fluorescence, or colorimetric methods.
