CDK9/CyclinK, GST-tag Recombinant (Discontinued)

Catalog #

40106

$410 *

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Description

Recombinant human full-length CDK9 (cyclin-dependent kinase) and full-length cyclin K. This complex contains an N-terminal GST-tag. The complex was affinity purified and is active.

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Synonyms

Cyclin-Dependent 9, CDK9: CDC2L4, C-2k, TAK, PITALRE
CyclinK: CCNK, CPR4, MGC9113, CDK-9

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

CDK9 (GST-Full Length) / CyclinK (GST-Full Length)

Host Species/Expression System

Sf9 insect cells

Purity

≥90%

Format

Aqueous buffer solution

Formulation

50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM Glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol.

68 kDa (CDK9), 67 kDa (CyclinK)

Amino Acids

full length

Specific Activity

3.0 pmol/min/µg

Genbank #

CDK9: NM_001261; CyclinK: NM_003858

UniProt #

CDK9: P50750; CyclinK: O75909

Tag(s)

N-terminal GST-tag

Background

CDK9/CyclinK is a member of the cyclin-dependent protein kinase (CDK) family. CDK9 is closely related to cdc28 and cdc2 and is an important regulator of the cell cycle (1). CDK9 is a component of the multiprotein complex TAK/P-TEFβ. CDK9 can modulate RNA polymerase II-directed transcription by phosphorylating the C-terminal domain of the largest subunit of RNA polymerase II. CDK9 forms a complex with and is regulated by its regulatory subunit cyclin T or cyclin K. CDK9 also interacts with the HIV-1 Tat protein which suggested a possible involvement of this protein in AIDS (2).

References

1. Yang, Z. et al: The 7SK small nuclear RNA inhibits the CDK9/cyclin T1 kinase to control transcription. Nature, 2001; 414: 317-322.
2. Bullrich, F. et al: Chromosomal mapping of members of the cdc2 family of protein kinases, cdk3, cdk6, PISSLRE, and PITALRE, and a cdk inhibitor, p27-Kip1, to regions involved in human cancer. Cancer Res. 1995; 55: 1199-1205.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

CDK9/Cyclin K activity was measured by using the PDKStide synthetic peptide (KTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC) diluted in 40 mM Tris-HCl (pH 7.5) to a working concentration of 1 mg/ml, in the ADP Glo™ Kinase Assay kit (Promega #V9101). Reaction was initiated by mixing increasing amounts of CDK9/Cyclin K with 25 μM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA prepared with 50 μM DTT and substrate at a final concentration of 200 µg/ml.

After a 40-minute incubation at room temperature, the reaction was terminated by addition of ADP-Glo™ Reagent, followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was added, and the reaction was incubated for another 30 minutes at ambient temperature. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate Multimode Reader. The Specific Activity was calculated as follows: (Corrected activity, RLU) / [(Specific activity from ADP in RLU/pmol) * (Reaction time in min) *(Enzyme amount in μg or mg)]. Corrected RLU was calculated by subtracting the blank value from all the values. The blank was determined from a “no enzyme” sample by replacing the enzyme solution with an equal volume of Dilution Buffer IX (1x).

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Kinase/Serine-Threonine

Regulatory Status

Research Use Only

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Datasheets/Protocols

40106 Lot 230823

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CDK9/CyclinK, GST-tag Recombinant (Discontinued)

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