Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade
| Name | Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab5408 |
| Prices | $404.00 |
| Sizes | 100 µg |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG1 |
| Clone | 4H8 |
| Applications | FC ICC/IF ICC/IF ChIP ChIP ChIP WB ELISA IP ChIPseq |
| Species Reactivities | Mouse, Rat, Human, Yeast, Arabidopsis thaliana, C. elegans, Drosophila, S. pombe, Monkey, Hamster |
| Antigen | Synthetic peptide: YSPTSpPS (Human) |
| Description | Mouse Monoclonal |
| Gene | CELE_F36A4.7 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
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.
![All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human RNA polymerase II CTD repeat YSPTSPS peptide (ab17564) at 1 µg/mlLane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488) at 1 µg/mlLane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with S. cerevisiae RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide (ab12793) at 1 µg/mlLysates/proteins at 20 µg/ml per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutionPerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/23558_RNA-polymerase-II-CTD-repeat-YSPTSPS-Primary-antibodies-ab5408-18.jpg)
All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (ab5408) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human RNA polymerase II CTD repeat YSPTSPS peptide (ab17564) at 1 µg/mlLane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488) at 1 µg/mlLane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with S. cerevisiae RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide (ab12793) at 1 µg/mlLysates/proteins at 20 µg/ml per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutionPerformed under reducing conditions.
HeLa or MCF7 cells were fixed with 4% formaldehyde in PEM buffer. The coverslip was incubated in blocking buffer of 5% powdered milk in TBS-T plus 0.02% sodium azide for 1 hour at room temperature. Blocking buffer was removed and primary antibody was added at a dilution of 1/1000 and incubated overnight at 4 degrees celsius. The coverslips were then washed 4-5 times with blocking buffer for 5 minutes. Secondary antibody, goat anti-mouse Alexa 594, was added at a dilution of 1/1000 and incubated at room temperature for one hour. From this point on coverslips were covered with foil to protect them from light. They were washed 5 times with TBS-T and then one time with PEM, for 5 minutes each wash. The coverslips were fixed 10-30 minutes in 4% formaldehyde in PEM buffer, then washed 3 times with PEM buffer for 5 minutes. 0.1M ammonium chloride in PEM buffer was added for 10 minutes to quench auto-florescence, and then slips were washed 2 times for 5 minutes in PEM followed by 3 washes for
ELISA using ab5408 at varying antibody concentrations. Curve_SPL4 indicates binding to RNA polymerase II CTD repeat YSPTSPS peptide - phospho S5 (ab18488). Binding to the following peptides was much weaker: Curve_SPL5 RNA polymerase II CTD repeat YSPTSPS peptide - phospho S2 (ab12793), Curve_SPL6 RNA polymerase II CTD repeat YSPTSPS peptide (ab12795).
![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.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/23562_RNA-polymerase-II-CTD-repeat-YSPTSPS-Primary-antibodies-ab5408-32.jpg)
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.
Product References
The Arabidopsis mediator complex subunits MED16, MED14, and MED2 regulate - The Arabidopsis mediator complex subunits MED16, MED14, and MED2 regulate
Hemsley PA, Hurst CH, Kaliyadasa E, Lamb R, Knight MR, De Cothi EA, Steele JF, Knight H. Plant Cell. 2014 Jan;26(1):465-84.
Canonical nucleosome organization at promoters forms during genome activation. - Canonical nucleosome organization at promoters forms during genome activation.
Zhang Y, Vastenhouw NL, Feng J, Fu K, Wang C, Ge Y, Pauli A, van Hummelen P, Schier AF, Liu XS. Genome Res. 2014 Feb;24(2):260-6.
A human artificial chromosome recapitulates the metabolism of native telomeres in - A human artificial chromosome recapitulates the metabolism of native telomeres in
Wakai M, Abe S, Kazuki Y, Oshimura M, Ishikawa F. PLoS One. 2014 Feb 18;9(2):e88530.
The up-regulation of miR-199b-5p in erythroid differentiation is associated with - The up-regulation of miR-199b-5p in erythroid differentiation is associated with
Li Y, Bai H, Zhang Z, Li W, Dong L, Wei X, Ma Y, Zhang J, Yu J, Sun G, Wang F. Mol Cells. 2014 Mar;37(3):213-9.
Therapeutic targeting of BET bromodomain proteins in castration-resistant - Therapeutic targeting of BET bromodomain proteins in castration-resistant
Asangani IA, Dommeti VL, Wang X, Malik R, Cieslik M, Yang R, Escara-Wilke J, Wilder-Romans K, Dhanireddy S, Engelke C, Iyer MK, Jing X, Wu YM, Cao X, Qin ZS, Wang S, Feng FY, Chinnaiyan AM. Nature. 2014 Jun 12;510(7504):278-82.
Antisense-mediated FLC transcriptional repression requires the P-TEFb - Antisense-mediated FLC transcriptional repression requires the P-TEFb
Wang ZW, Wu Z, Raitskin O, Sun Q, Dean C. Proc Natl Acad Sci U S A. 2014 May 20;111(20):7468-73. doi:
EFNA3 long noncoding RNAs induced by hypoxia promote metastatic dissemination. - EFNA3 long noncoding RNAs induced by hypoxia promote metastatic dissemination.
Gomez-Maldonado L, Tiana M, Roche O, Prado-Cabrero A, Jensen L, Fernandez-Barral A, Guijarro-Munoz I, Favaro E, Moreno-Bueno G, Sanz L, Aragones J, Harris A, Volpert O, Jimenez B, Del Peso L. Oncogene. 2014 Jul 14;0.
Pre-mRNA splicing is a determinant of nucleosome organization. - Pre-mRNA splicing is a determinant of nucleosome organization.
Keren-Shaul H, Lev-Maor G, Ast G. PLoS One. 2013;8(1):e53506.
RNA-interference components are dispensable for transcriptional silencing of the - RNA-interference components are dispensable for transcriptional silencing of the
Cernilogar FM, Burroughs AM, Lanzuolo C, Breiling A, Imhof A, Orlando V. PLoS One. 2013 Jun 13;8(6):e65740.
Mediator complex recruits epigenetic regulators via its two cyclin-dependent - Mediator complex recruits epigenetic regulators via its two cyclin-dependent
Tsutsui T, Fukasawa R, Shinmyouzu K, Nakagawa R, Tobe K, Tanaka A, Ohkuma Y. J Biol Chem. 2013 Jul 19;288(29):20955-65.