Western Blot

graph TD A[Cell lysate] --> Lyse[Lyse cells in RIPA buffer] B[Loading buffer] --> AND1{Lysate AND
Loading buffer?} Lyse --> AND1 AND1 --> Prep[Prepare samples] Prep --> Load[Load onto gel] Load --> PAGE[SDS-PAGE electrophoresis] PAGE --> Equil[Equilibrate gel in transfer buffer] Equil --> Transfer[Transfer to PVDF/nitrocellulose] Transfer --> Block[Block with BSA or milk] Block --> PAb[Incubate primary antibody] PAb --> Wash1[Wash membrane] Wash1 --> SAb[Incubate HRP-conjugated secondary Ab] SAb --> Wash2[Wash membrane] Wash2 --> Sub[Add chemiluminescent substrate] Sub --> Image[Expose to film or imager] style A fill:#ff6b6b,stroke:#c92a2a,color:#fff style B fill:#ff6b6b,stroke:#c92a2a,color:#fff style Lyse fill:#51cf66,stroke:#2f9e44,color:#fff style Prep fill:#51cf66,stroke:#2f9e44,color:#fff style Load fill:#51cf66,stroke:#2f9e44,color:#fff style PAGE fill:#51cf66,stroke:#2f9e44,color:#fff style Equil fill:#74c0fc,stroke:#1971c2,color:#fff style Transfer fill:#51cf66,stroke:#2f9e44,color:#fff style Block fill:#51cf66,stroke:#2f9e44,color:#fff style PAb fill:#51cf66,stroke:#2f9e44,color:#fff style Wash1 fill:#51cf66,stroke:#2f9e44,color:#fff style SAb fill:#51cf66,stroke:#2f9e44,color:#fff style Wash2 fill:#51cf66,stroke:#2f9e44,color:#fff style Sub fill:#51cf66,stroke:#2f9e44,color:#fff style Image fill:#b197fc,stroke:#7950f2,color:#fff style AND1 fill:#b4b4dc,stroke:#8b8bc4,color:#000