Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575) is a rabbit monoclonal antibody that is used to detect RNA polymerase II CTD repeat YSPTSPS in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF, ChIP, Dot Blot. Suitable for Human, Mouse, Rat samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency
Images
![Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--western-blot-img431309.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575)")
![Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--western-blot-img400607.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575)")
![Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunoprecipitation-img400757.jpg/_jcr_content/renditions/webp-475.webp "Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575)")
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img400658.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575)")
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img400660.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (AB300575)")
Publications
Filter by
- Science advances 9:eadd50052023Nucleic DHX9 cooperates with STAT1 to transcribe interferon-stimulated genes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXingxing Ren,Decai Wang,Guorong Zhang,Tingyue Zhou,Zheng Wei,Yi Yang,Yunjiang Zheng,Xuqiu Lei,Wanyin Tao,Anmin Wang,Mingsong Li,Richard A Flavell,Shu ZhuPubMed 36735791
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| ChIP | IP | Flow Cyt (Intra) | ICC/IF | IHC-P | Dot | WB | |
|---|---|---|---|---|---|---|---|
| Human | Tested | Tested | Tested | Tested | Tested | Expected | Tested |
| Mouse | Expected | Not recommended | Tested | Tested | Tested | Expected | Tested |
| Rat | Expected | Expected | Expected | Expected | Tested | Expected | Tested |
| Synthetic peptide - Human | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended | Tested | Not recommended |
ChIP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 5 µg/mL | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1/30 | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse, Synthetic peptide - Human | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species Mouse | Dilution info 1/500 | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1/50 | Notes - |
Species Human | Dilution info 1/50 | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info 1/5000 | Notes Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Mouse | Dilution info 1/5000 | Notes Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/5000 | Notes Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
Dot
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide - Human | Dilution info 1/1000 | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1/5000 | Notes - |
Species Rat | Dilution info 1/5000 | Notes - |
Species Human | Dilution info 1/5000 | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
Target data
Function
Catalytic core component of RNA polymerase II (Pol II), a DNA-dependent RNA polymerase which synthesizes mRNA precursors and many functional non-coding RNAs using the four ribonucleoside triphosphates as substrates (By similarity) (PubMed:23748380, PubMed:27193682, PubMed:30190596, PubMed:9852112). Pol II-mediated transcription cycle proceeds through transcription initiation, transcription elongation and transcription termination stages. During transcription initiation, Pol II pre-initiation complex (PIC) is recruited to DNA promoters, with focused-type promoters containing either the initiator (Inr) element, or the TATA-box found in cell-type specific genes and dispersed-type promoters that often contain hypomethylated CpG islands usually found in housekeeping genes. Once the polymerase has escaped from the promoter it enters the elongation phase during which RNA is actively polymerized, based on complementarity with the template DNA strand. Transcription termination involves the release of the RNA transcript and polymerase from the DNA (By similarity) (PubMed:23748380, PubMed:27193682, PubMed:28108474, PubMed:30190596, PubMed:9852112). Forms Pol II active center together with the second largest subunit POLR2B/RPB2. Appends one nucleotide at a time to the 3' end of the nascent RNA, with POLR2A/RPB1 most likely contributing a Mg(2+)-coordinating DxDGD motif, and POLR2B/RPB2 participating in the coordination of a second Mg(2+) ion and providing lysine residues believed to facilitate Watson-Crick base pairing between the incoming nucleotide and template base. Typically, Mg(2+) ions direct a 5' nucleoside triphosphate to form a phosphodiester bond with the 3' hydroxyl of the preceding nucleotide of the nascent RNA, with the elimination of pyrophosphate. The reversible pyrophosphorolysis can occur at high pyrophosphate concentrations (By similarity) (PubMed:30190596, PubMed:8381534, PubMed:9852112). Can proofread the nascent RNA transcript by means of a 3' -> 5' exonuclease activity. If a ribonucleotide is mis-incorporated, backtracks along the template DNA and cleaves the phosphodiester bond releasing the mis-incorporated 5'-ribonucleotide (By similarity) (PubMed:8381534). Through its unique C-terminal domain (CTD, 52 heptapeptide tandem repeats) serves as a platform for assembly of factors that regulate transcription initiation, elongation and termination. CTD phosphorylation on Ser-5 mediates Pol II promoter escape, whereas phosphorylation on Ser-2 is required for Pol II pause release during transcription elongation and further pre-mRNA processing. Additionally, the regulation of gene expression levels depends on the balance between methylation and acetylation levels of the CTD-lysines. Initiation or early elongation steps of transcription of growth-factor-induced immediate early genes are regulated by the acetylation status of the CTD. Methylation and dimethylation have a repressive effect on target genes expression. Cooperates with mRNA splicing machinery in co-transcriptional 5'-end capping and co-transcriptional splicing of pre-mRNA (By similarity) (PubMed:24207025, PubMed:26124092). RNA-dependent RNA polymerase that catalyzes the extension of a non-coding RNA (ncRNA) at the 3'-end using the four ribonucleoside triphosphates as substrates. An internal ncRNA sequence near the 3'-end serves as a template in a single-round Pol II-mediated RNA polymerization reaction. May decrease the stability of ncRNAs that repress Pol II-mediated gene transcription. (Microbial infection) Acts as an RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicase and transcriptase for the viral RNA circular genome.
Alternative names
POLR2, POLR2A, DNA-directed RNA polymerase II subunit RPB1, RNA polymerase II subunit B1, 3'-5' exoribonuclease, DNA-directed RNA polymerase II subunit A, DNA-directed RNA polymerase III largest subunit, RNA-directed RNA polymerase II subunit RPB1
Recommended products
Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575) is a rabbit monoclonal antibody that is used to detect RNA polymerase II CTD repeat YSPTSPS in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF, ChIP, Dot Blot. Suitable for Human, Mouse, Rat samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Clone number
- EPR24494-59
- Purification technique
- Affinity purification Protein A
- Specificity
This antibody is unsuitable for mouse IP.
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage duration
- 1-2 weeks
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- -20°C
- Aliquoting information
- Upon delivery aliquot
- Storage information
- Avoid freeze / thaw cycle
Notes
Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), ChIP, Dot Blot, in Human, Mouse, Rat samples.
Recommended positive controls
WB: Whole cell lysates: HeLa (human cervix adenocarcinoma epithelial cell), 293T (human embryonic kidney epithelial cell), NIH/3T3 (mouse embryonic fibroblast), PC-12 (rat adrenal gland pheochromocytoma), RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage). IHC-P: Human, mouse, and rat colon. ICC/IF: HeLa, RAW 264.7. Flow cyt. Intra.: HeLa, RAW 264.7. IP: HeLa. ChIP: Hela. DB: RNA polymerase II CTD repeat YSPTSPS non-phosphorylated and (phospho Y1/S7) peptide.
Trial sizes available!
Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.Other related products
We have a range of other formats of antibody clone [EPR24494-59] also available for your convenience:
ab300575, Carrier free - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] - ChIP Grade - BSA and Azide free ab300576, PE - PE Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] – ChIP Grade ab318393, APC - APC Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] – ChIP Grade ab318496, Alexa Fluor® 488 - Alexa Fluor® 488 Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] – ChIP Grade ab318599, Alexa Fluor® 647 - Alexa Fluor® 647 Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] – ChIP Grade ab318702, Alexa Fluor® 594 - Alexa Fluor® 594 Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] – ChIP Grade ab318805, Alexa Fluor® 555 - Alexa Fluor® 555 Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] – ChIP Grade ab318905, Alexa Fluor® 750 - Alexa Fluor® 750 Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] - ChIP Grade ab321548
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free batch production
For more information, read more on recombinant antibodies.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
RNA polymerase II CTD repeat YSPTSPS also known as the C-terminal domain of RNA polymerase II is a critical component of the RNA polymerase II enzyme commonly referred to as pol II. This domain is characterized by the repetitive sequence YSPTSPS which plays a significant role in the regulation of transcription. The mass of RNA polymerase II including its CTD varies but is essential for its function in gene expression. RNA polymerase II with the CTD is expressed in the nucleus of eukaryotic cells where it orchestrates the transcription of DNA into mRNA.
- Biological function summary
RNA polymerase II CTD repeat YSPTSPS is essential for the transcription progression from initiation to termination. It is part of the large RNA polymerase II complex interacting with various transcription factors and enzymes necessary for RNA processing. The phosphorylation state of the CTD particularly on serine residues regulates interactions with splicing machinery and other RNA processing factors. This modulation ensures the coupling between transcription and RNA processing events controlling mRNA synthesis and maturation.
- Pathways
RNA polymerase II CTD repeat YSPTSPS is important in the mRNA synthesis pathway specifically in transcriptional regulation and processing of nascent RNA transcripts. It interacts with proteins such as the transcription factors TFIIH and TFIIB which aid in promoter recognition and open complex formation. The CTD's dynamic phosphorylation pattern allows integration into multiple cellular pathways most importantly connecting transcription with RNA splicing and transport pathways.
- Associated diseases and disorders
Abnormal function or mutations in RNA polymerase II CTD repeat YSPTSPS associate with diseases such as transcription-related syndromes and certain cancers. Deficient CTD phosphorylation can lead to improper mRNA processing resulting in neural developmental disorders. Additionally its interaction with proteins like CDK7 which phosphorylates the CTD links it to tumors where transcriptional dysregulation is a hallmark. Understanding the CTD's role in these diseases provides insight into therapeutic targets and strategies for intervention.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
10 product images
![Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--western-blot-img431309.jpg "Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
RNA polymerase II CTD repeat YSPTSPS Western blot staining using rabbit Anti-RNA polymerase II CTD repeat YSPTSPS antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] staining at ChIP Grade dilution.
All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [RM1152] (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [RM1152] ab317456) at 1/1000 dilution
Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg
Lane 2: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate (untreated membrane) at 20 µg
Lane 3: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate (untreated membrane) at 20 µg
Lane 4: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate (untreated membrane) at 20 µg
Lane 5: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 6: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 7: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
Lane 8: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 270 kDa, 124 kDa
Exposure time: 15s
![Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--western-blot-img400607.jpg "Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
All lanes: Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575) at 1/5000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 5: RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 250 kDa
Exposure time: 3s
![Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunoprecipitation-img400757.jpg "Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
RNA polymerase II CTD repeat YSPTSPS was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 2 ug with ab300575 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300575 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 2 ug
Lane 2: abAB300575 IP in HeLa whole cell lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab300575 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 secondsAll lanes: Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575) at 1/1000 dilution
All lanes: HeLa whole cell lysate
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 10s
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img400658.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling RNA polymerase II CTD repeat YSPTSPS with ab300575 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used. Nuclear staining on human colon (PMID: 25901683). The section was incubated with ab300575 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img400660.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling RNA polymerase II CTD repeat YSPTSPS with ab300575 at 1/5000 (0.108 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used. Nuclear staining on rat colon. The section was incubated with ab300575 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img421695.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling RNA polymerase II CTD repeat YSPTSPS with ab300575 at 1/5000 dilution (0.108 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Nuclear staining on mouse colon is observed. The section was incubated with ab300575 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.![Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunocytochemistry-immunofluorescence-img400725.jpg "Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling RNA polymerase II CTD repeat YSPTSPS with ab300575 at 1/50 (10.78 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing mainly nuclear staining in HeLa cell line is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.
![Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunocytochemistry-immunofluorescence-img400726.jpg "Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling RNA polymerase II CTD repeat YSPTSPS with ab300575 at 1/50 (10.78 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing mainly nuclear staining in RAW 264.7 cell line is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/mL dilution.
![ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--chip-img426028.jpg "ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
Chromatin was prepared from HeLa cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 5 µg of ab300575 (red), or 5 µg of rabbit normal IgG Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.
![Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--dot-blot-img426027.jpg "Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)
Dot blot analysis of RNA polymerase II CTD repeat YSPTSPS using ab300575 at 1:1000 (0.539 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.
Lane 1: RNA polymerase II CTD repeat YSPTSPS (phospho Y1) peptide
Lane 2: RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide
Lane 3: RNA polymerase II CTD repeat YSPTSPS (phospho T4) peptide
Lane 4: RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide
Lane 5: RNA polymerase II CTD repeat YSPTSPS (phospho S7) peptide
Lane 6: RNA polymerase II CTD repeat YSPTSPS non-phopho peptide
Exposure time: 3 minutes.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img421695.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
![Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunocytochemistry-immunofluorescence-img400725.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
![Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--immunocytochemistry-immunofluorescence-img400726.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
![ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--chip-img426028.jpg/_jcr_content/renditions/webp-475.webp "ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
![Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575), expandable thumbnail](https://content.abcam.com/products/images/rna-polymerase-ii-ctd-repeat-ysptsps-antibody-epr24494-59-chip-grade-ab300575--dot-blot-img426027.jpg/_jcr_content/renditions/webp-475.webp "Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS antibody [EPR24494-59] (ChIP Grade) (ab300575)")
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