Cdk1 cyclin b

Manufactured by Sino Biological

CDK1/cyclin B is a protein complex that plays a crucial role in cell cycle regulation. It is composed of the catalytic subunit CDK1 (cyclin-dependent kinase 1) and the regulatory subunit cyclin B. This complex is responsible for promoting the transition from G2 phase to mitosis (M phase) during the cell cycle.

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Lab products found in correlation

Kinase buffer, by New England Biolabs (2 mentions) Gstrap ff affinity columns, by GE Healthcare (2 mentions) Recombinant cdk1 cyclin b complex, by New England Biolabs (2 mentions) γ 32p atp, by PerkinElmer (2 mentions)

2 protocols using cdk1 cyclin b

Phosphorylation Assay for GST-PBK

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GST-tagged PBK and PBK-4A were cloned in pGEX-5X-1 and proteins were bacterially expressed and purified on GSTrap FF affinity columns (GE Healthcare) following the manufacturer’s instructions. GST-PBK (0.5–1 μg) was incubated with 10 U recombinant CDK1/cyclin B complex (New England Biolabs) or 100 ng CDK1/cyclin B (SignalChem) in kinase buffer (New England Biolabs) in the presence of 5 μCi γ-³²P-ATP (3000 Ci/mmol, PerkinElmer). Phosphorylation (³²P incorporation) was visualized by autoradiography followed by Western blotting or detected by phospho-specific antibodies.

Stauffer S., Zeng Y., Zhou J., Chen X., Chen Y, & Dong J. (2017). CDK1-mediated Mitotic Phosphorylation of PBK is Involved in Cytokinesis and Inhibits its Oncogenic Activity. Cellular signalling, 39, 74-83.

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In Vitro Phosphorylation of MST2

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GST-tagged MST2 or MST2-S385A (cloned in pGEX-5X-1) was bacterially expressed and purified on GSTrap FF affinity columns (GE Healthcare) following the manufacturer’s instructions. GST-MST2 (1 μg) was incubated with 5-10 U recombinant CDK1/cyclin B complex (New England Biolabs) or 50-100 ng CDK1/cyclin B (SignalChem) in kinase buffer (New England Biolabs) in the presence of 5 μCi γ-³²P-ATP (3000 Ci/mmol, PerkinElmer) as we previously described [15 (link)]. Active CDK2, CDK5, p38, JNK1, JNK2, MEK1, ERK1, and PLK1 kinases were also purchased from SignalChem.

Chen X., Chen Y, & Dong J. (2016). MST2 Phosphorylation at Serine 385 in Mitosis Inhibits its Tumor Suppressing Activity. Cellular signalling, 28(12), 1826-1832.

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