Anti-CDC27 pT244 Antibody - Western Blot. Western blot of Affinity Purified anti-CDC27 pT244 antibody shows detection of a band ~92 kD corresponding to phosphorylated human CDC27 (arrowhead). Lane 1 shows lysate from asynchronous cells. Lane 2 shows lysate from cells treated with nocodazole. Phosphorylated CDC27 is mostly present only in cell preparations arrested in mitosis. Each lane contains approximately 30 ug of HeLa whole cell lysates separated by 12.5% SDS-PAGE followed by transfer to nitrocellulose. After blocking with 5% non-fat dry milk in TTBS, the membrane was probed with the primary antibody diluted to 1:500 for 1 h at room temperature followed by washes and reaction with a 1:5000 dilution of HRP Gt-a-Rabbit IgG [H&L] MX (LS-C60865) for 45 min at room temperature. ECL reagent was used for detection. Other detection systems will yield similar results. Data contributed by Bing Li, UT Southwestern.