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06-1289
Sigma-AldrichAnti-Cyclin T1 Antibody
Use Anti-Cyclin T1 Antibody (Sheep Polyclonal Antibody) validated in WB to detect Cyclin T1 also known as CDK9-associated C-type protein, cyclin C-related protein.
More>>Use Anti-Cyclin T1 Antibody (Sheep Polyclonal Antibody) validated in WB to detect Cyclin T1 also known as CDK9-associated C-type protein, cyclin C-related protein. Less<<
Anti-Cyclin T1 Antibody: SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
human immunodeficiency virus type 1 (HIV-1) expression (elevated) 1
subunit of positive elongation transcription factor b
Background Information
Cyclin T1, which is also known as CCNT, is a member of the cyclin family and functions as a CDK kinase regulator. Cyclin T1 is one of regulatory cyclins (T1, T2a, T2b, and K) that is closely associated with CDK9 kinase forming a heterodimer called P-TEFb. This kinase complex is involved in phosporylating the carboxyl-terminal domain (CTD) of the large RNA polymerase II subunit which stimulate the elongation upon initiation. The cyclin T1/CDK9 complex has also been referred to as tat-associated kinase. Tat is an HIV protein that targets the cyclin T1/CDK9 complex to activate the transcription of the virus.
Use Anti-Cyclin T1 Antibody (Sheep Polyclonal Antibody) validated in WB to detect Cyclin T1 also known as CDK9-associated C-type protein, cyclin C-related protein.
Key Applications
Western Blotting
Application Notes
Western Blot Analysis: A 1:1000 dilution from a previous lot detected Cyclin T1 in 10 µg of HEK293, HeLa, HepG2, Jurkat, and PC3 cell lysates.
Biological Information
Immunogen
His-tagged recombinant protein corresponding to human Cyclin T1.
Epitope
Unknown
Host
Sheep
Specificity
This antibody recognizes Cyclin T1.
Isotype
IgG
Species Reactivity
Human
Mouse
Species Reactivity Note
Proven to react with human and mouse. Rat (85% sequence homology).
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]
FUNCTION: Regulatory subunit of the cyclin-dependent kinase pair (CDK9/cyclin-T1) complex, also called positive transcription elongation factor B (P-TEFb), which is proposed to facilitate the transition from abortive to productive elongation by phosphorylating the CTD (carboxy-terminal domain) of the large subunit of RNA polymerase II (RNA Pol II). In case of HIV or SIV infections, binds to the transactivation domain of the viral nuclear transcriptional activator, Tat, thereby increasing Tat's affinity for the transactivating response RNA element (TAR RNA). Serves as an essential cofactor for Tat, by promoting RNA Pol II activation, allowing transcription of viral genes.
SUBUNIT STRUCTURE: Cyclin-T1 is the predominant cyclin that associates with CDK9 to form a heterodimer called P-TEFb. P-TEFb forms a complex with AFF4/AF5Q31. Interacts with the transactivation region of HIV-1, HIV-2 and SIV Tat. Component of a complex which is at least composed of HTATSF1/Tat-SF1, P-TEFb complex, RNA pol II, SUPT5H, and NCL/nucleolin. Component of the 7SK snRNP complex at least composed of P-TEFb (composed of CDK9 and CCNT1/cyclin-T1), HEXIM1, HEXIM2, BCDIN3, SART3 proteins and 7SK and U6 snRNAs. Interacts with MDFIC.
SUBCELLULAR LOCATION: Nucleus.
TISSUE SPECIFICITY: Ubiquitously expressed.
MISCELLANEOUS: Interaction between Tat and cyclin-T1 requires zinc.
SEQUENCE SIMILARITIES: Belongs to the cyclin family. Cyclin C subfamily.
Molecular Weight
Cyclin T1 is expected to be ~85 kDa. This antibody detects a ~85 kDa band in most cell lines. It also detects the degraded forms of Cyclin T1 (~70 kDa, ~60 kDa, et al), which is beneficial for IP applications. According to independent laboratory, this antibody can immunoprecipitate multiple forms of Cyclin T1.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Western Blot in HeLa cell lysate.
Western Blot Analysis: A 1:1000 dilution of this antibody detected Cyclin T1 in 10 µg of HeLa cell lysate.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
The mechanism of release of P-TEFb and HEXIM1 from the 7SK snRNP by viral and cellular activators includes a conformational change in 7SK. Krueger, BJ; Varzavand, K; Cooper, JJ; Price, DH PloS one
5
e12335
2009
The positive transcription elongation factor, P-TEFb, is required for the production of mRNAs, however the majority of the factor is present in the 7SK snRNP where it is inactivated by HEXIM1. Expression of HIV-1 Tat leads to release of P-TEFb and HEXIM1 from the 7SK snRNP in vivo, but the release mechanisms are unclear.We developed an in vitro P-TEFb release assay in which the 7SK snRNP immunoprecipitated from HeLa cell lysates using antibodies to LARP7 was incubated with potential release factors. We found that P-TEFb was directly released from the 7SK snRNP by HIV-1 Tat or the P-TEFb binding region of the cellular activator Brd4. Glycerol gradient sedimentation analysis was used to demonstrate that the same Brd4 protein transfected into HeLa cells caused the release of P-TEFb and HEXIM1 from the 7SK snRNP in vivo. Although HEXIM1 binds tightly to 7SK RNA in vitro, release of P-TEFb from the 7SK snRNP is accompanied by the loss of HEXIM1. Using a chemical modification method, we determined that concomitant with the release of HEXIM1, 7SK underwent a major conformational change that blocks re-association of HEXIM1.Given that promoter proximally paused polymerases are present on most human genes, understanding how activators recruit P-TEFb to those genes is critical. Our findings reveal that the two tested activators can extract P-TEFb from the 7SK snRNP. Importantly, we found that after P-TEFb is extracted a dramatic conformational change occurred in 7SK concomitant with the ejection of HEXIM1. Based on our findings, we hypothesize that reincorporation of HEXIM1 into the 7SK snRNP is likely the regulated step of reassembly of the 7SK snRNP containing P-TEFb.
