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Synthetic peptide corresponding to Human RNA polymerase II CTD repeat YSPTSPS (phospho S5). The sequence is repeated multiple times in the C-terminal domain of RNA polymerase II.
Database link: P24928
Our Abpromise guarantee covers the use of ab5408 in the following tested applications.
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ChIP||Use 1-4µg for 106 cells.|
|ChIP/Chip||Use at an assay dependent concentration. PubMed: 19703992|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 260 kDa (predicted molecular weight: 217 kDa).|
|ELISA||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration. See Abreviews.|
|CHIPseq||Use 2-0.3 µg for µg of chromatin.|
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab5408 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
Lane 1: buffer.
Lane 2: extract from human 293T cells as positive control.
Lanes 3 and 4: egg extracts made from wild type N2 worms and worms grown on RNAi food against dicer.
Lanes 5-8: intact worms in mentioned larval stage and eggs boiled directly in SDS-PAGE buffer and loaded.
This image was kindly supplied as part of the review submitted by Shveta Bagga.
Overlay histogram showing HeLa cells stained with ab5408 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5408, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.