Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5)抗体
参阅全部 RNA polymerase II CTD repeat YSPTSPS 一抗
兔多克隆抗体to RNA polymerase II CTD repeat YSPTSPS (phospho S5)
Rabbit
This antibody recognises the phosphorylated serine found in the amino acid 5 position of the C-terminal domain repeat YSPTSPS of RNA polymerase II. From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help.
适用于: IHC-Fr, IHC-P, ELISA, IP, WB, ChIP, ICC/IF, IHC - Wholemountmore details
与反应: Mouse, Rat, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Zebrafish
预测可用于: a wide range of other species
This product was produced with the following immunogens:
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab18488)
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab18488)
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab18488)
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab18488)
ICC/IF: HeLa cells. WB: Xenopus laevis whole tissue lysate
Phosphorylation of RNA polymerase II's largest subunit C-terminal domain (CTD) is a key event during mRNA metabolism.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
浓度
批次浓度范围 50 µg 浓度为 0.6 - 1 mg/ml
Immunogen affinity purified
Phosphorylation of RNA polymerase II's largest subunit C-terminal domain (CTD) is a key event during mRNA metabolism.
多克隆
IgG
Abpromise™承诺保证使用ab5131于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| IHC-Fr | Use at an assay dependent concentration. | |
| IHC-P | Use at an assay dependent concentration. | |
| ELISA | (1) | Use at an assay dependent concentration. |
| IP | (1) | Use at an assay dependent concentration. |
| WB | (15) | 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 | (17) | Use 2 µg for 25 µg of chromatin. |
| ICC/IF | (9) | Use a concentration of 1 µg/ml. |
| IHC - Wholemount | (2) | Use at an assay dependent concentration. |
Entrez Gene: 177190 Caenorhabditis elegans
Entrez Gene: 32100 Drosophila melanogaster
Entrez Gene: 5430 Human
Entrez Gene: 20020 Mouse
Entrez Gene: 851415 Saccharomyces cerevisiae
Omim: 180660 Human
SwissProt: P18616 Arabidopsis thaliana
SwissProt: P16356 Caenorhabditis elegans
SwissProt: P04052 Drosophila melanogaster
SwissProt: P24928 Human
SwissProt: P08775 Mouse
SwissProt: P04050 Saccharomyces cerevisiae
Unigene: 2925 Drosophila melanogaster
Unigene: 270017 Human
Unigene: 16533 Mouse
DNA directed RNA polymerase II A antibody
DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit antibody
DNA-directed RNA polymerase II subunit A antibody
DNA-directed RNA polymerase II subunit RPB1 antibody
DNA-directed RNA polymerase III largest subunit antibody
hRPB220 antibody
hsRPB1 antibody
POLR2 antibody
Polr2a antibody
POLRA antibody
Polymerase (RNA) II (DNA directed) polypeptide A 220kDa antibody
Polymerase (RNA) II (DNA directed) polypeptide A antibody
RNA polymerase II subunit B1 antibody
RNA-directed RNA polymerase II subunit RPB1 antibody
RPB1 antibody
RPB1_HUMAN antibody
RPBh1 antibody
RpIILS antibody
RPO2 antibody
RPOL2 antibody
ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody - ChIP Grade (ab5131)Murgatroyd et al PLoS One. 2014 Mar 5;9(3):e90277. doi: 10.1371/journal.pone.0090277. eCollection 2014. Fig S2. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Chromatin from cultured cells, mouse PVN punches (individual pools formed from groups of 3) or whole hypothalami dissected from fresh brains were cross linked, disrupted by sonification and purified.
The binding of Suz12 to wingless (Wnt1), a beta-catenin dependent developmental regulator and to the RNA polymerase II promoter (RNAPII), a housekeeping gene, served as positive and negative controls, respectively, in these experiments.
ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody - ChIP Grade (ab5131)
Chromatin was prepared from U-2 OS (Human bone osteosarcoma epithelial cell line) cells according to the Abcam X-ChIP protocol.
Cells were fixed with formaldehyde for 10 minutes. 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.
Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab5131)
ab5131 staining RNA polymerase II CTD repeat YSPTSPS (phospho S5) in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab5131 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody - ChIP Grade (ab5131)This image is courtesy of an anonymous Abreview
All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab5131) at 1/5000 dilution
Lane 1 : Xenopus laevis whole tissue lysate treated with DMSO for 24 hours
Lane 2 : Xenopus laevis whole tissue lysate treated with CDK inhibitor for 24 hours
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 217 kDa
Observed band size: 240 kDawhy is the actual band size different from the predicted?
Exposure time: 1 minute
Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody - ChIP Grade (ab5131)
ICC/IF image of ab5131 stained HeLa(Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed in methanol for 5 minutes, permabilized in TBS-T for 20 minutes and incubated with the antibody (ab5131, 1 µg/ml) for 1 hour at room temperature. 1% BSA / 10% normal goat serum / 0.3M glycine was used to quench auto-fluorescence and block non-specific protein-protein interactions.
The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody - ChIP Grade (ab5131)
All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab5131) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 2 : S.cerevisiae (Y190) Whole Cell Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate with S. cerevisiae RNA polymerase II CTD repeat YSPTSPS peptide (ab12795) at 1 µg/ml
Lane 4 : S.cerevisiae (Y190) Whole Cell Lysate with S. cerevisiae RNA polymerase II CTD repeat YSPTSPS peptide (ab12795) at 1 µg/ml
Lane 5 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate with Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488) at 1 µg/ml
Lane 6 : S.cerevisiae (Y190) Whole Cell Lysate with Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488) at 1 µg/ml
Lane 7 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate with S. cerevisiae RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide (ab12793) at 1 µg/ml
Lane 8 : S.cerevisiae (Y190) Whole Cell Lysate with S. cerevisiae RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide (ab12793) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 217 kDa
Exposure time: 30 seconds
Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody - ChIP Grade (ab5131)
Lanes 1 & 3-4 : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab5131) at 1/500 dilution
Lane 2 : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody (ab5131) at 1/2000 dilution
Lanes 1-2 : HeLa nuclear extract
Lane 3 : HeLa nuclear extract with Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide (ab18488) at 1 µg/ml
Lane 4 : HeLa nuclear extract with S. cerevisiae RNA polymerase II CTD repeat YSPTSPS peptide (ab12795) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 217 kDa
Observed band size: 250 kDawhy is the actual band size different from the predicted?
Exposure time: 30 seconds
