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06-1291
Sigma-AldrichAnti-Hexim1 Antibody
Anti-Hexim1 Antibody is a Sheep polyclonal antibody for detection of Hexim1 also known as Cardiac lineage protein 1, Estrogen down-regulated gene 1 protein ,Hexamethylene bis-acetamide-inducible protein 1 & has been validated in WB.
More>>Anti-Hexim1 Antibody is a Sheep polyclonal antibody for detection of Hexim1 also known as Cardiac lineage protein 1, Estrogen down-regulated gene 1 protein ,Hexamethylene bis-acetamide-inducible protein 1 & has been validated in WB. Less<<
Anti-Hexim1 Antibody: SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
Hexim1 is a transcriptional regulator which functions as a general RNA polymerase II transcription inhibitor. Hexim1 works in cooperation with 7SK snRNA to sequester P-TEFb in a large inactive 7SK snRNP complex, preventing RNA polymerase II phosphorylation and subsequent transcriptional elongation. Hexim1 may also regulate NF-kappa-B, ESR1, NR3C1 and CIITA-dependent transcriptional activity. The 7SK snRNP complex is composed of P-TEFb (composed of CDK9 and CCNT1/cyclin-T1), Hexim1, Hexim2, BCDIN3, SART3 proteins and 7SK and U6 snRNAs. Hexim1 also interacts with the N-CoR complex through NCOR1, ESR1, NR3C1, and may interact with NF-kappa-B through REL A. Hexim1 is located in the nucleus where it can bind with alpha-importin, and can also be found in the cytoplasm. Hexim1 is up-regulated by HMBA (hexamethylene bisacetamide) at the protein level and down-regulated by estrogen.
Anti-Hexim1 Antibody is a Sheep polyclonal antibody for detection of Hexim1 also known as Cardiac lineage protein 1, Estrogen down-regulated gene 1 protein ,Hexamethylene bis-acetamide-inducible protein 1 & has been validated in WB.
Key Applications
Western Blotting
Application Notes
Western Blot Analysis: A 1:1,000 dilution from a previous lot detected Hexim1 in 10 µg of HeLa cell lysate.
Biological Information
Immunogen
His-tagged recombinant protein corresponding to human Hexim1.
Host
Sheep
Specificity
This antibody recognizes Hexim1.
Species Reactivity
Human
Species Reactivity Note
Proven to react with human. Mouse (85% sequence homology) and rat (86% sequence homology).
FUNCTION: Transcriptional regulator which functions as a general RNA polymerase II transcription inhibitor. In cooperation with 7SK snRNA sequesters P-TEFb in a large inactive 7SK snRNP complex preventing RNA polymerase II phosphorylation and subsequent transcriptional elongation. May also regulate NF-kappa-B, ESR1, NR3C1 and CIITA-dependent transcriptional activity. SUBUNIT STRUCTURE: Homooligomer and heterooligomer with HEXIM2; probably dimeric. 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 the N-CoR complex through NCOR1. Interacts with ESR1 and NR3C1. May interact with NF-kappa-B through RELA. SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Note: Binds alpha-importin and is mostly nuclear. TISSUE SPECIFICITY: Ubiquitously expressed with higher expression in placenta. HEXIM1 and HEXIM2 are differentially expressed. Expressed in endocrine tissues. INDUCTION: Up-regulated by HMBA (hexamethylene bisacetamide) (at protein level). Down-regulated by estrogen. DOMAIN: The coiled-coil domain mediates oligomerization. MISCELLANEOUS: Inhibits Tat activity which is required for HIV-1 transcription. SEQUENCE SIMILARITIES: Belongs to the HEXIM family.
Molecular Weight
~40 kDa (predicted). Molecular weight Note: According to an independent laboratory, the two isoforms of Hexim1 run at ~53 kDa & ~68 kDa. This antibody can detect the ~53 kDa isoform in multiple cells.
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 1:1,000 dilution of this antibody detected Hexim1 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 first thaw, 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.