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Synthetic peptide of Saccharomyces cerevisiae RNA polymerase II CTD repeat YSPTSPS, phosphorylated at S5.
(Peptide available as ab18488.)
Phosphorylation of RNA polymerase II's largest subunit C-terminal domain (CTD) is a key event during mRNA metabolism.
Our Abpromise guarantee covers the use of ab5131 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC - Wholemount||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|CHIPseq||Use at an assay dependent concentration.|
|WB||1/1000. Detects a band of approximately 240 kDa (predicted molecular weight: 217 kDa).Can be blocked with Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488).|
|ChIP||Use 2-25 µg for µg of chromatin.|
|ICC/IF||Use a concentration of 1 µg/ml.|
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 mins. The ChIP was performed with 25 µg of chromatin, 2 µg of ab5131 (blue), and 20 µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the inactive AFM and F8 promoters, the GAPDH promoter (active) and over the y-Actin gene (active). Schematic diagram of the y-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.
IHC - Wholemount of Caenorhabditis elegans larvae labelling RNA polymerase II CTD repeat YSPTSPS (phospho S5) with ab5131. The sample was incubated with primary antibody (1/500 in PBS + 3% BSA + 0.1% Triton X-100) for 12 hours at 4°C. ab150077, a goat anti-rabbit Alexa 488 - (1/1000), was used as the secondary antibody.
Western blot using ab5131.
Lane 1: ab5131 at 1/500
Lane 2: ab5131 at 1/2000
Lane 3: ab5131 at 1/500 blocked with phospho peptide
Lane 4: ab5131 at 1/500 blocked with non-phospho control peptide
Phospho peptide is YSPTSpPSYSPTSpPS-GGC (ab18488)
Non-phospho control peptide is YSPTSPSYSPTSPS-GGC (ab12795)
Secondary ab: Goat anti-rabbit HRP (IgG) ab6721 (1/5000)
Lanes 1 to 4: 20µg of HeLa nuclear extract per lane
Blocking peptides used at 1µg/ml.
Exposure time: 30 seconds
Expected molecular weight: ~240 kDa
This image is courtesy of an anonymous Abreview
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"