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AB264109

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free

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(1 Publication)

Mouse Recombinant Monoclonal RNA polymerase II RPB1 phospho S5 antibody. Carrier free. Suitable for Flow Cyt (Intra), ICC/IF, ChIP, ELISA, Dot, WB and reacts with Mouse, Human, Rat, Synthetic peptide - Human samples. Cited in 1 publication.

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

12 Images
Flow Cytometry - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • Flow Cyt

Unknown

Flow Cytometry - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

Flow cytometry overlay histogram showing HeLa cells stained with ab264109 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab264109) (1x106 in 100 μL at 0.2 μg/ml) for 30 min at 22°C.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 22°C.

Isotype control antibody (black line) was mouse IgG1κ; (ab170190) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in HeLa cells fixed with 4 % formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized HeLa cells with ab5408 at 1/500 dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used for counterstain at a 1/100 dilution (Red). Nuclear DNA was labelled with DAPI (shown in blue)

Confocal image showing nuclear staining in HeLa cell line, the signal decreased after phosphatase treatment at 37°C for 2h.

Flow Cytometry (Intracellular) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S5) with ab5408 at 1/1000 dilution (0.1ug) followed by Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) secondary at 1/2000 dilution (Red) compared with a Mouse monoclonal IgG isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • IP

Lab

Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

RNA polymerase II CTD repeat YSPTSPS (phospho S5) was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab5408 at 1/50 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab5408 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (<a href='/en-us/products/primary-antibodies/rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-antibody-4h8-chip-grade-ab5408'>ab5408</a>) at 1/50 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

HeLa whole cell lysate

Lane 3:

Mouse IgG1, kappa monoclonal isotype control (ab184443) instead of <a href='/en-us/products/primary-antibodies/rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-antibody-4h8-chip-grade-ab5408'>ab5408</a> in HeLa whole cell lysate

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 240 kDa

false

Exposure time: 10s

ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • ChIP

Lab

ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab5408 (red), or 5 µg of Mouse IgG1 ab18443 (gray) and 25 µl of Protein A/G Dynabeads. 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.
*http : //www.abcam.com/resources?keywords=X%20ChIP%20protocol

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells with ab5408 at 1/500 dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used for counterstain at a 1/100 dilution (Red). Nuclear DNA was labelled with DAPI (shown in blue)

Confocal image showing nuclear staining in NIH/3T3 cell line, the signal decreased after phosphatase treatment at 37°C for 2h.

Flow Cytometry (Intracellular) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S5) with ab5408 at 1/1000 dilution (0.1ug) followed by Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) secondary at 1/2000 dilution (Red) compared with a Mouse monoclonal IgG isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • WB

Lab

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

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 : 23071310)

All lanes:

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (<a href='/en-us/products/primary-antibodies/rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-antibody-4h8-chip-grade-ab5408'>ab5408</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate (Untreated membrane) at 10 µg

Lane 2:

MCF7 whole cell lysate (Phosphatase treated membrane) at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Predicted band size: 260 kDa

false

Exposure time: 3.25s

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • WB

Lab

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

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 : 23071310)

All lanes:

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (<a href='/en-us/products/primary-antibodies/rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-antibody-4h8-chip-grade-ab5408'>ab5408</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Predicted band size: 260 kDa

false

Exposure time: 15s

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • WB

Lab

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

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 : 23071310)

All lanes:

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (<a href='/en-us/products/primary-antibodies/rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-antibody-4h8-chip-grade-ab5408'>ab5408</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate (Untreated membrane) at 10 µg

Lane 2:

NIH/3T3 whole cell lysate (Phosphatase treated membrane) at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Predicted band size: 260 kDa

false

Exposure time: 3.25s

Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • I-ELISA

Lab

Indirect ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

Indirect ELISA analysis of ab5408 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Mouse IgG (H+L) at 1/1000 dilution.

Antigen : RNA polymerase II CTD (phospho S2) peptide, phospho S5 peptide, phospho T4 peptide, phospho Y1peptide, phospho S7 peptide, non-phospho peptide.

Antigen concentration : 100 ng/ml

Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)
  • Dot

Lab

Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - BSA and Azide free (AB264109)

This image was produced using the same antibody clone but in a different formulation, ab5408.

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

All lanes:

Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade (<a href='/en-us/products/primary-antibodies/rna-polymerase-ii-ctd-repeat-ysptsps-phospho-s5-antibody-4h8-chip-grade-ab5408'>ab5408</a>) at 1/1000 dilution

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

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Predicted band size: 260 kDa

false

Exposure time: 3min

  • Unconjugated

    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [4H8] - ChIP Grade

Key facts

Host species

Mouse

Clonality

Monoclonal

Clone number

4H8

Isotype

IgG1

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

ChIP, Dot, ELISA, ICC/IF, WB, Flow Cyt (Intra)

applications

Immunogen

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

style
left-column

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "ChIP" : {"fullname" : "ChIP", "shortname":"ChIP"}, "ELISA" : {"fullname" : "ELISA", "shortname":"ELISA"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/mouse-igg1-kappa-monoclonal-15-6e10a7-isotype-control-ab170190'>ab170190</a> - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.</p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "ChIP-species-checked": "testedAndGuaranteed", "ChIP-species-dilution-info": "", "ChIP-species-notes": "<p></p>", "ELISA-species-checked": "guaranteed", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "Mouse": { "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "ChIP-species-checked": "guaranteed", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "guaranteed", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "Rat": { "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "guaranteed", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "guaranteed", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "African green monkey": { "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "predicted", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "predicted", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "predicted", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Arabidopsis thaliana": { "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "predicted", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "predicted", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "predicted", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Caenorhabditis elegans": { "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "predicted", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "predicted", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "predicted", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Hamster": { "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "predicted", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "predicted", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "predicted", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Monkey": { "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "predicted", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "predicted", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "predicted", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Saccharomyces cerevisiae": { "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "predicted", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "predicted", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "predicted", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Schizosaccharomyces pombe": { "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "predicted", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "predicted", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "Dot-species-checked": "predicted", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Synthetic peptide - Human": { "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "ChIP-species-checked": "notRecommended", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "ELISA-species-checked": "testedAndGuaranteed", "ELISA-species-dilution-info": "", "ELISA-species-notes": "<p></p>", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "" } } }
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Product details

ab264109 is the carrier-free version of ab5408.

ChIP protocols:
ChIP protocol for cross-linking ChIP (X-ChIP)
Native ChIP protocol
Chromatin preparation from tissues for ChIP
ChIP troubleshooting
ChIP tips and tricks guide

Want a custom formulation?
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

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.

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Properties and storage information

Form
Liquid
Purity
IgG fraction
Storage buffer
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
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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.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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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
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Publications (1)

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

The Journal of cell biology 200:61-80 PubMed23277425

2013

Neurobeachin regulates neurotransmitter receptor trafficking to synapses.

Applications

Unspecified application

Species

Unspecified reactive species

Ramya Nair,Juliane Lauks,SangYong Jung,Nancy E Cooke,Heidi de Wit,Nils Brose,Manfred W Kilimann,Matthijs Verhage,JeongSeop Rhee
View all publications
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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.
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