CDK4-CCND2 Protein, Human (Active, sf9, GST, Flag, His)

CDK4 protein, acting as a Ser/Thr kinase in the cyclin D-CDK4 complex, critically regulates the G(1)/S transition by phosphorylating and inhibiting RB family members. This activity, specifically hypophosphorylation of RB1 during early G(1) phase, releases E2F and promotes transcription of genes that drive G(1) progression. CDK4-CCND2 Protein, Human (sf9, GST, Flag, His) is the recombinant human-derived CDK4-CCND2, expressed by Sf9 insect cells, with N-His, N-GST, N-Flag labeled tag.

CDK4 protein serves as the Ser/Thr-kinase component within cyclin D-CDK4 (DC) complexes, orchestrating the phosphorylation and inhibition of members belonging to the retinoblastoma (RB) protein family, including RB1. This regulatory activity is pivotal in controlling the cell cycle during the G(1)/S transition. The phosphorylation of RB1 instigates the dissociation of the transcription factor E2F from the RB/E2F complexes, facilitating the subsequent transcription of E2F target genes that drive progression through the G(1) phase. Particularly notable is the hypophosphorylation of RB1 occurring in early G(1) phase. As essential integrators of diverse mitogenic and antimitogenic signals, cyclin D-CDK4 complexes play a central role in cell cycle regulation. Additionally, CDK4 protein exhibits the capability to phosphorylate SMAD3 in a cell-cycle-dependent manner, thereby repressing its transcriptional activity. CDK4 is a crucial component of the ternary complex, cyclin D/CDK4/CDKN1B, which is indispensable for the nuclear translocation and activity of the cyclin D-CDK4 complex.

The activity was measured by off-chip mobility shift assay(MSA). The enzyme was incubated with fluorecence-labeled substrate and Mg(or Mn)/ATP. The phosphorylated and unphosphorylated substrates were separated and detected by MSA device. The Km of CDK4-CCND2 for its substrate is 165 μM.

Human

Sf9 insect cells

N-His;N-GST;N-Flag

P11802-1 (M1-E303)&P30279-1 (E2-L289)

1019&894

60.3 KDa & 36.0 KDa

≥ 85%, as determined by reducing SDS-PAGE.

Solution.

Supplied as a 0.2 μm filtered solution of 50 mM HEPES (pH 7.5), 200 mM NaCl, 5% glycerol.

<1 EU/μg, determined by LAL method.

Please use rapid thawing with running water to thaw the protein.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.