Active Motif

Phospho-RNA pol II CTD (Ser5) Monoclonal Antibody (3E8)

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Phospho-RNA pol II CTD (Ser5) Antibody in ChIP assay (ChIP) — RNA pol II CTD phospho Ser5 antibody (mAb) tested by ChIP. Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 10 µg of chromatin from human myeloma LP1 cells and 10 µg RNA pol II CTD phospho Ser5 antibody. ChIP DNA was used in qPCR with the control primer pairs or gene-specific primer pairs as indicated. Data are presented as Binding Events Detected per 1000 Cells using Active Motifs Epigenetic Services normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

Phospho-RNA pol II CTD (Ser5) Antibody in ChIP-sequencing (ChIP-Seq) — RNA pol II CTD phospho Ser5 antibody (mAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with chromatin from 2.3 million HL60 cells and 5 ug of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 23 million sequence tags were mapped to identify RNA pol II phospho Ser5 binding. Data is compared to ChIP-Seq data using a phospho Ser2 antibody (61083). ChIP-Seq data from three specific genes is shown as an example. The Pol II phospho Ser5 antibody detects polymerase more at the 5180 end of the genes and the phospho Ser2 antibody detects Pol II more toward the 3180 end of the genes.

Product Details

61085

Applications	Tested Dilution	Publications
Western Blot (WB)	0.5-2 µg/mL	-
Immunocytochemistry (ICC/IF)	1:500	-
Immunoprecipitation (IP)	-	View 1 publication 1 publication
ChIP assay (ChIP)	5-10 µg	View 3 publications 3 publications
ChIP-sequencing (ChIP-Seq)	5-10 µg	View 2 publications 2 publications
Miscellaneous PubMed (Misc)	-	View 4 publications 4 publications

Product Specifications
Species Reactivity	Human
Published species	Human, Rat, Yeast
Host/Isotype	Rat / IgG2a
Class	Monoclonal
Type	Antibody
Clone	3E8
Immunogen	This RNA pol II CTD phospho Ser5 antibody was raised against a peptide containing the RNA pol II CTD sequence phosphorylated at serine 5. View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein G
Storage buffer	70mM tris, pH 8, with 30% glycerol, 31mM glycine, 105mM NaCl, 0.07mM EDTA
Contains	0.035% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2687451

Target Information

RNA polymerase II is the enzyme responsible for the synthesis of mRNA and many non-coding RNAs. POLR2A encodes RNA polymerase II subunit B1 (RPB1), the largest subunit of RNA polymerase II. The C-terminal domain (CTD) of RPB1 contains tandem heptapeptide repeats with the consensus sequence YSPTSPS. Phosphorylation and other modifications within this sequence influence the recruitment of factors that regulate the transcription cycle. Recruitment of RNA processing factors to the elongation complex is coordinated by the phosphorylation of Serine-2 and Serine-5 residues within the CTD.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Amyloid-like Assembly Activates a Phosphatase in the Developing Drosophila Embryo.

Cell

Nil Z,Hervás R,Gerbich T,Leal P,Yu Z,Saraf A,Sardiu M,Lange JJ,Yi K,Unruh J,Slaughter B,Si K

Research topic: Embryology, Developmental Biology

Thu Sep 05 00:00:00 EDT 2019

Ring finger protein 121 is a potent regulator of adeno-associated viral genome transcription.

PLoS pathogens

Madigan VJ,Yuziuk JA,Chiarella AM,Tyson TO,Meganck RM,Elmore ZC,Tse LV,Hathaway NA,Asokan A

Research topic: Transcription

Thu Aug 01 00:00:00 EDT 2019

Histone Hypervariants H2A.Z.1 and H2A.Z.2 Play Independent and Context-Specific Roles in Neuronal Activity-Induced Transcription of Arc/Arg3.1 and Other Immediate Early Genes.

eNeuro

Dunn CJ,Sarkar P,Bailey ER,Farris S,Zhao M,Ward JM,Dudek SM,Saha RN

Research topic: Brain & Nerves

Mon Sep 17 00:00:00 EDT 2018

NF-κB inhibition rescues cardiac function by remodeling calcium genes in a Duchenne muscular dystrophy model.

Nature communications

Peterson JM,Wang DJ,Shettigar V,Roof SR,Canan BD,Bakkar N,Shintaku J,Gu JM,Little SC,Ratnam NM,Londhe P,Lu L,Gaw CE,Petrosino JM,Liyanarachchi S,Wang H,Janssen PML,Davis JP,Ziolo MT,Sharma SM,Guttridge DC

Research topic: Transcription

Fri Aug 24 00:00:00 EDT 2018

Real-time observation of light-controlled transcription in living cells.

Journal of cell science

Rademacher A,Erdel F,Trojanowski J,Schumacher S,Rippe K

Research topic: Gene Regulation

Fri Dec 15 00:00:00 EST 2017

Acetylation on histone H3 lysine 9 mediates a switch from transcription initiation to elongation.

The Journal of biological chemistry

Gates LA,Shi J,Rohira AD,Feng Q,Zhu B,Bedford MT,Sagum CA,Jung SY,Qin J,Tsai MJ,Tsai SY,Li W,Foulds CE,O'Malley BW

Fri Sep 01 00:00:00 EDT 2017

Comprehensive RNA Polymerase II Interactomes Reveal Distinct and Varied Roles for Each Phospho-CTD Residue.

Cell reports

Harlen KM,Trotta KL,Smith EE,Mosaheb MM,Fuchs SM,Churchman LS

Research topic: Transcription Regulation

Tue Jun 07 00:00:00 EDT 2016

DNA demethylation by 5-aza-2'-deoxycytidine is imprinted, targeted to euchromatin, and has limited transcriptional consequences.

Epigenetics & chromatin

Ramos MP,Wijetunga NA,McLellan AS,Suzuki M,Greally JM

Research topic: Transcription Regulation

Wed Mar 25 00:00:00 EDT 2015

Cyclin-dependent kinase 12 increases 3' end processing of growth factor-induced c-FOS transcripts.

Molecular and cellular biology

Eifler TT,Shao W,Bartholomeeusen K,Fujinaga K,Jäger S,Johnson JR,Luo Z,Krogan NJ,Peterlin BM

Research topic: Transcription Regulation

Thu Jan 01 00:00:00 EST 2015

Chromatin and transcription transitions of mammalian adult germline stem cells and spermatogenesis.

Cell stem cell

Hammoud SS,Low DH,Yi C,Carrell DT,Guccione E,Cairns BR

Research topic: Spermatogenesis, Stem Cells

Thu Aug 07 00:00:00 EDT 2014

Bioinformatics

Protein Aliases: DNA-directed RNA polymerase II largest subunit, RNA polymerase II 220 kd subunit; DNA-directed RNA polymerase II subunit A; DNA-directed RNA polymerase II subunit RPB1; DNA-directed RNA polymerase III largest subunit; EC 2.7.7.6; MGC75453; Pol II S2p; polymerase (RNA) II (DNA directed) polypeptide A, 220kDa; polymerase (RNA) II subunit A; POLYMERASE II; RNA; RNA POL2; RNA polymerase II subunit B1; RNA-directed RNA polymerase II subunit RPB1; RP02; RPB220; RPO21; SUA8

Gene Aliases: hRPB220; hsRPB1; POLR2; POLR2A; POLRA; RPB1; RPBh1; RpIILS; RPO2; RPOL2

UniProt ID: (Human) P24928

Entrez Gene ID: (Human) 5430

Molecular Function: DNA-directed RNA polymerase