In mammals, cyclin B associates with inactive p34 cdc2 which facilitates phosphorylation of p34 cdc2 at amino acids 14 Thr and 15 Tyr. This maintains the inactive state until the end of G2 phase. The inactive cyclin B p34 cdc2 complex continues to accumulate in the cytoplasm until the completion of DNA synthesis, when Cdc25, a specific protein phosphatase, dephosphorylates amino acids 14Thr and 15Tyr of p34 cdc2, 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. Therefore, cyclin B p34 cdc2 plays a critical role in G2 to M transition. Two alternative transcripts have been found, a constitutively expressed transcript, and a cell cycle-regulated transcript that is expressed predominantly during G2/M phase. The different transcripts result from the use of alternate transcription initiation sites.
This antibody recognizes a Ser126 phosphorylation involved in an auto phosphorylation of the cdk -cyclin complex. Recognition of this phosphorylated site suggests the antibody is useful to detect the active form of the complex and may serve as a marker of mitosis.
Applications:
Suitable for use in ELISA and Western Blot. Other applications not tested.
Recommended Dilution:
ELISA: 1:10,000-1:50,000
Western Blot: 1:100-1:1000
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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