AB255846

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free

Recombinant
What is this?

Be the first to review this product! Submit a review

(2 Publications)

Rat Recombinant Monoclonal RNA polymerase II RPB1 phospho S5 antibody. Carrier free. Suitable for ICC/IF, ChIP, Dot, WB, I-ELISA and reacts with Mouse, Human, Rat, Synthetic peptide samples. Cited in 2 publications.

View 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

8 Images

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S5) with ab252852 at 1/500 (1.29 μg/mL) dilution , followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2 dilution (Green). Confocal image showing nuclear staining in HeLa cell line, the signal decreased after phosphatase treatment at 37°C for 2h. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/500 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

ChIP

Lab

ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

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 10min.
The ChIP was performed with 25 μg of chromatin, 5 μg of ab252852 (red), or 5 μg of Rat IgG2a ab18450 (gray) and 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW264.7 cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S5) with ab252852 at 1/500 (1.29 μg/mL) dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in RAW264.7 cell line, the signal decreased after phosphatase treatment at 37°C for 2h. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) secondary antibody at 1/500 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

Lab

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22745433 and 23071310).

Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

All lanes:

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - ChIP Grade (<a href='/en-us/products/primary-antibodies/rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-antibody-3e8-chip-grade-ab252852'>ab252852</a>) at 0.645 µg/mL

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate (Untreated membrane) at 10 µg

Lane 2:

HeLa whole cell lysate (Phosphatase treated membrane) at 10 µg

Secondary

All lanes:

Goat Anti-Rat IgG (H+L), HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 217 kDa

Observed band size: 260 kDa

false

Lab

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22745433 and 23071310).

Exposure time : 37 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

All lanes:

Lane 1:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate (Untreated membrane) at 10 µg

Lane 2:

RAW264.7 whole cell lysate (Phosphatase treated membrane) at 10 µg

Secondary

All lanes:

Goat Anti-Rat IgG (H+L), HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 217 kDa

Observed band size: 260 kDa

false

Lab

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22745433 and 23071310).

Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

All lanes:

Lane 1:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 2:

PC-12 whole cell lysate (Phosphatase treated membrane) at 10 µg

Secondary

All lanes:

Goat Anti-Rat IgG (H+L), HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 217 kDa

Observed band size: 260 kDa

false

Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

I-ELISA

Lab

Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

ab252852 was used at 0 - 1000 ng/mL.

Antigens were used at 1000 ng/mL.

An Alkaline Phosphatase-conjugated Anti-Rat IgG (H+L) wasused as secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

Lab

Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

RNA polymerase II CTD repeat YSPTSPS (phospho S5) labeled with ab252852 at 1/1000 (0.645 μg/mL) dilution.

Goat Anti-Rat IgG (H+L), HRP) (ab205720) at 1/5000 dilution was used as secondary antibody.

Lane 1 : RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide

Lane 2 : RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide

Lane 3 : RNA polymerase II CTD repeat YSPTSPS (phospho T4) peptide

Lane 4 : RNA polymerase II CTD repeat YSPTSPS (phospho Y1) peptide

Lane 5 : RNA polymerase II CTD repeat YSPTSPS (phospho S7) peptide

Lane 6 : RNA polymerase II CTD repeat YSPTSPS non-phopho peptide

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

Unconjugated

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - ChIP Grade

Key facts

Host species

Rat

Clonality

Monoclonal

Clone number

3E8

Isotype

IgG2a

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

WB, I-ELISA, Dot, ChIP, ICC/IF

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

style

left-column

style

left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "ChIP" : {"fullname" : "ChIP", "shortname":"ChIP"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IELISA" : {"fullname" : "Indirect ELISA", "shortname":"I-ELISA"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "ChIP-species-checked": "testedAndGuaranteed", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IELISA-species-checked": "guaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" }, "Mouse": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "ChIP-species-checked": "guaranteed", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IELISA-species-checked": "guaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" }, "Rat": { "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "ChIP-species-checked": "guaranteed", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IELISA-species-checked": "guaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" }, "Synthetic peptide": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "ChIP-species-checked": "notRecommended", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IELISA-species-checked": "testedAndGuaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" } } }

style

left-column

Product details

ab255846 is the carrier-free version of ab252852.

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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

style

left-column

Properties and storage information

Form

Liquid

Purification technique

Ion exchange chromatography

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

style

left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.

style

left-column

style

left-column

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

style

left-column

Target data

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.

See full target information POLR2A phospho S5

style

left-column

Publications (2)

Recent publications for all applications. Explore the full list and refine your search

Transcription 6:91-101 PubMed26566685

2015

Site-specific methylation and acetylation of lysine residues in the C-terminal domain (CTD) of RNA polymerase II.

Applications

Unspecified application

Species

Unspecified reactive species

Kirsten Voss,Ignasi Forné,Nicolas Descostes,Corinna Hintermair,Roland Schüller,Muhammad Ahmad Maqbool,Martin Heidemann,Andrew Flatley,Axel Imhof,Marta Gut,Ivo Gut,Elisabeth Kremmer,Jean-Christophe Andrau,Dirk Eick

Science (New York, N.Y.) 318:1780-2 PubMed18079404

2007

Transcribing RNA polymerase II is phosphorylated at CTD residue serine-7.

Applications

Unspecified application

Species

Unspecified reactive species

Rob D Chapman,Martin Heidemann,Thomas K Albert,Reinhard Mailhammer,Andrew Flatley,Michael Meisterernst,Elisabeth Kremmer,Dirk Eick

View all publications

style

left-column

style

left-column

style

right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (AB255846)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Complementary Products

Primary Antibodies

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (2)

Product promise