Recombinant CDK4 / Cyclin D1 Complex

Catalog No: 82084 Format: 20 µg $480 Add to Cart
Catalog No: 82684 Format: 1 mg $4,150 Add to Cart

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Expressed In: Baculovirus Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

Cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G1/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G1 phase. Hypophosphorylates RB1 in early G1 phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex.

Application Notes

This product was manufactured as described in Protein Details. Where possible, Active Motif has developed functional or activity assays for recombinant proteins. Additional characterization such as enzyme kinetic activity assays, inhibitor screening or other biological activity assays may not have been performed for every product.  All available data for this product is shown. 

Protein Details

Recombinant CDK4 / Cyclin D1 Complex that includes full-length human CDK4 protein (accession number NP_000066.1) with an N-terminal His tag and full-length human Cyclin D1 protein (accession number NP_444284.1) with an C-terminal FLAG tag was expressed in Hi5 cells. The molecular weights of CDK4 and Cyclin D1 are 36.83 kDa and 35.42 kDa, respectively.

References

Human

 

Recombinant CDK4/Cyclin D1 protein gel
10% SDS-PAGE Coomassie staining
MW: CDK4: 36.83 kDa, Cyclin D1: 35.42 kDa
Purity: >90%

ADP-Glo assay for CDK4 / Cyclin D1 Complex activity
0.02 µg/µl Rb (773 - 928) was incubated with different concentrations of CDK4 / Cyclin D1 Complex protein in a 5 µl reaction system containing 40 mM Tris-HCl pH 7.4, 20 mM MgCl2, 1 mM DTT, 0.1 mg/ml BSA and 10 µM ATP at 30℃ for 1 hour, then 5 µl ADP-Glo Reagent was added. After 40 minutes of incubation at RT, 10 µl Kinase Detection Reagent was added in the system. Following 30 minutes incubation at RT, luminescence was detected with a plate reader.

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is guaranteed for 6 months from date of receipt. This product is for research use only and is not for use in diagnostic procedures.