Product Name :
EAF1 polyclonal antibody Background :
Actin is a key regulator of RNA polymerase (Pol) II-dependent transcription. Positive transcription elongation factor b (P-TEFb), a Cdk9/cyclin T1 heterodimer, has been reported to play a critical role in transcription elongation. However, the relationship between actin and P-TEFb is still not clear. In this study, actin was found to interact with Cdk9, a catalytic subunit of P-TEFb, in elongation complexes. Using immunofluorescence and immunoprecipitation assays, Cdk9 was found to bind to G-actin through the conserved Thr-186 in the T-loop. Overexpression and in vitro kinase assays showed that G-actin promotes P-TEFb-dependent phosphorylation of the Pol II C-terminal domain. An in vitro transcription experiment revealed that the interaction between G-actin and Cdk9 stimulated Pol II transcription elongation. ChIP and immobilized template assays indicated that actin recruited Cdk9 to a transcriptional template in vivo and in vitro. Using cytokine IL-6-inducible p21 gene expression system, we revealed that actin recruited Cdk9 to endogenous gene. Moreover, overexpression of actin and Cdk9 increased histone H3 acetylation and acetylized histone H3 binding to a transcriptional template through the interaction with histone acetyltransferase, p300. Taken together, our results suggested that actin participates in transcription elongation by recruiting Cdk9 for phosphorylation of the Pol II C-terminal domain, and the actin-Cdk9 interaction promotes chromatin remodeling. Product :
1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2 Storage&Stability :
Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles. Specificity :
Unmodification Immunogen :
Recombinant fusion protein of human EAF1(NP_149074.3). Conjugate :
Unconjugated Modification :
Unmodification

• 
  Western blot analysis of extracts of various cell lines, using EAF1 antibody at 1:1000 dilution.
  Secondary antibody: HRP Goat Anti-Rabbit IgG at 1:10000 dilution.
  Lysates/proteins: 25ug per lane.
  Blocking buffer: 3% nonfat dry milk in TBST.
  Detection: ECL Basic Kit .
  Exposure time: 60s.

Bioworld Biotech only provide peptides for our antibodies and do not provide additional peptide customization services.

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Price/Size :

USD 368/1mg/vial

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Tips: 

For phospho antibody, we provide phospho peptide（0.5mg) and non-phospho peptide(0.5mg).

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Describe :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (WB) and Immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or IHC performed with the neutralized antibody.

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Formula:

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.

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Storage:

The freeze-dried powder is more stable. For short time at 2-8°C. For long term storage store at -20°C. 

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Note :

This product is for research use only (RUO only). Not for use in diagnostic or therapeutic procedures.

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