EAF1 Polyclonal Antibody-BS77490

EAF1 Polyclonal Antibody

Sizes: 50µl, 100µl

Catalogue Numbers: BS77490-50, BS77490-100

Product: 1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2

Swiss-Prot: Q96JC9

Host: Rabbit

Reactivity: Human, Rat

Applications: WB

All Applications: WB,1:500 - 1:2000

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.

Purification and Purity: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).

Storage and Stability: Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.

Specificity: Unmodification

Extra Notes: 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.

Note: For research use only, not for use in diagnostic procedure.

Bioworld Molecular Weight: 29kDa

Alternative Name: EAF1

Immunogen: Recombinant fusion protein of human EAF1(NP_149074.3).

Conjugate: Unconjugated

Modification: Unmodified