Total cell lysates (15 µg protein) from HepG2 (Lane 1), HeLa (Lane 2), Raji (Lane 3), HEL (Lane 4), or RAW264.7 (Lane 5) cells were resolved using 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with purified anti-Cbl antibody (Poly6330) at a 1:1000 dilution. Proteins were visualized by chemiluminescence detection using HRP Donkey anti-rabbit-IgG at a 1:3000 dilution. Equal protein loading was confirmed using Direct-Blot HRP anti-β-actin antibody at a 1:5000 dilution (lower). Lane M: MW ladder. Cell lysates were loaded in order of increasing CBL mRNA expression levels (source: Human Protein Atlas).