It recognizes a protein of 55-62kDa, identified as cyclin B1. In mammals, cyclin B associates with inactive p34cdc2, which facilitates phosphorylation of p34cdc2 at aa 14Thr and 15Tyr. This maintains the inactive state until the end of G2-phase. The inactive cyclin B-p34cdc2 complex continues to accumulate in the cytoplasm until the completion of DNA synthesis, when Cdc25, a specific protein phosphatase, dephosphorylates aa 14Thr and 15Tyr of p34cdc2 rendering the complex active at the G2/M boundary. This mitotic kinase complex remains active until the metaphase/anaphase transition when cyclin B is degraded. This degradation process is ubiquitin-dependent and is necessary for the cell to exit mitosis. So, cyclin B-p34cdc2 plays a critical role in G2 to M transition.
Flow Cytometry (1-2ug/million cells), Immunofluorescence (1-2ug/ml), Immunohistochemistry (Formalin-fixed) (1-2ug/ml for 30 min at RT)(Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95
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