Western blot analysis was performed on membrane extracts (30 µg lysate) of HeLa (Lane 1) and MCF-7 (Lane 2). The blots were probed with Anti-CDK5RAP2 Recombinant Rabbit Monoclonal Antibody (Product # 702394, 1-2 µg/mL). A 250 kDa band corresponding to CDK5RAP2 was observed as shown. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with wet transfer. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).