Western blot analysis of extracts of various cell lines, using EAF1 antibody (A17798) at 1:1000 dilution.|Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.|Lysates/proteins: 25ug per lane.|Blocking buffer: 3% nonfat dry milk in TBST.|Detection: ECL Basic Kit (RM00020).|Exposure time: 60s.
EAF1 Rabbit pAb
Catalog No:AB-A17798
Manufacturer:ABclonal
Synonyms:EAF1
Applications:WB
Reactivity:Human,Rat
Host:Rabbit
Conjugate:No
€65,00 – €370,00
Product Name: | EAF1 Rabbit pAb |
Category: | Polyclonal Antibodies |
Description: | 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. |
Immunogen: | Recombinant fusion protein containing a sequence corresponding to amino acids 50-120 of human EAF1 (NP_149074.3). |
Gene ID: | 85403 |
Swiss prot: | Q96JC9 |
Molecular weight: | 17kDa/29kDa |
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