Western blot analysis of extracts from 293 cells, using Phospho-Cyclin B1 (Ser147) Antibody (A) and Cyclin B1 (V152) Monoclonal Antibody (C). Treatment of the membrane with calf intestinal alkaline phosphatase (CIP) after Western transfer abolishes the phospho-cyclin B1 signal (B), but has no effect on the total cyclin B1 signal (D).
Western blot analysis of extracts from asynchronous cells (lane 1), S-phase and G2/M blocked cells (lane 2), or synchronized cells collected at various time points following release from this block (lanes 3-7) using Phospho-Cyclin B1 (Ser147) Antibody (upper). Cell cycle synchronization was verified by flow cytometric analysis of DNA content (lower). (Data kindly provided by Ethan Kohn and Alan Eastman, Dartmouth Medical School, Hanover, NH).