Human CDK9 (NP_001252.1, 1 a.a. - 372 a.a.) full-length recombinant protein, N-terminal GST-HIS6 fusion protein with a Thrombin cleavage site and CCNT1 (NP_001231.2, 1 a.a. - 726 a.a.) full-length recombinant protein, N-terminal HIS6 fusion protein with a Thrombin cleavage site co-expressed in Sf9 insect cells.
Host:
insect
Form:
Liquid
Preparation Method:
Insect cell (Sf9) expression system
Purification:
GST-Affinity chromatography
Concentration:
0.168 ug/uL
Activity:
Specific kinase activity (Pi transfer): 26 pmol/ug x min ATP-KM: 4 uM
Quality Control Testing:
2 ug/lane SDS-PAGE Stained with Coomassie Blue
Storage Buffer:
50 mM HEPES, 100 mM NaCl, pH 7.5 (5 mM DTT, 4 mM reduced glutathione, 20% glycerol).
Storage Instruction:
Store at -80°C. Aliquot to avoid repeated freezing and thawing. For complete recovery, mix well and spin before use. Product must not be stored in diluted solutions, aliquots below 10 uL are not advisable.
Note:
Result of activity analysis
Determination of Vmax and KM value for ATP
Assay conditions: 60 mM HEPES-NaOH, pH 7.5 3 mM MgCl2 3 mM MnCl2 3 uM Na-orthovanadate 1.2 mM DTT 50 ug/mL PEG20.000 ATP (variable) Substrate: RB-CTF, 200 ug/mL Kinase: 4 ug/mL
Filter binding assay MSFC membrane (Millipore)
Theoretical MW:
GST-CDK9: 72,173 Da HIS-CCNT1: 85,441 Da
Activation:
This kinase was not activated by special procedures.
The protein encoded by this gene is a member of the cyclin-dependent protein kinase (CDK) family. CDK family members are highly similar to the gene products of S. cerevisiae cdc28, and S. pombe cdc2, and known as important cell cycle regulators. This kinase was found to be a component of the multiprotein complex TAK/P-TEFb, which is an elongation factor for RNA polymerase II-directed transcription and functions by phosphorylating the C-terminal domain of the largest subunit of RNA polymerase II. This protein forms a complex with and is regulated by its regulatory subunit cyclin T or cyclin K. HIV-1 Tat protein was found to interact with this protein and cyclin T, which suggested a possible involvement of this protein in AIDS. [provided by RefSeq
Other Designations:
CDC2-related kinase,OTTHUMP00000022198,cell division protein kinase 9,serine/threonine protein kinase PITALRE
The protein encoded by this gene belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance through the cell cycle. Cyclins function as regulators of CDK kinases. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. This cyclin tightly associates with CDK9 kinase, and was found to be a major subunit of the transcription elongation factor p-TEFb. The kinase complex containing this cyclin and the elongation factor can interact with, and act as a cofactor of human immunodeficiency virus type 1 (HIV-1) Tat protein, and was shown to be both necessary and sufficient for full activation of viral transcription. This cyclin and its kinase partner were also found to be involved in the phosphorylation and regulation of the carboxy-terminal domain (CTD) of the largest RNA polymerase II subunit. [provided by RefSeq
Other Designations:
CDK9-associated C-type protein,cyclin C-related protein,cyclin T1b,subunit of positive elongation transcription factor b