Anti-POLR2A Antibody (CAB11181)

Antibody Name:	Anti-POLR2A Antibody
Antibody SKU:	CAB11181
Antibody Size:	20uL, 50uL, 100uL
Application:	WB IHC IF IP ChIP
Reactivity:	Human, Mouse, Rat
Host Species:	Rabbit
Immunogen:	Recombinant fusion protein containing a sequence corresponding to amino acids 1-260 of human POLR2A (NP_000928.1).

Application:	WB IHC IF IP ChIP
Recommended Dilution:	WB 1:500 - 1:2000 IHC 1:50 - 1:200 IF 1:50 - 1:200 IP 1:20 - 1:50 ChIP 1:20 - 1:50
Reactivity:	Human, Mouse, Rat
Positive Samples:	HeLa, BT-474, H460, Jurkat, Mouse testis, Mouse brain

Immunogen:	Recombinant fusion protein containing a sequence corresponding to amino acids 1-260 of human POLR2A (NP_000928.1).
Purification Method:	Affinity purification
Storage Buffer:	Store at -20'C. Avoid freeze / thaw cycles. Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.
Isotype:	IgG
Sequence:	MHGG GPPS GDSA CPLR TIKR VQFG VLSP DELK RMSV TEGG IKYP ETTE GGRP KLGG LMDP RQGV IERT GRCQ TCAG NMTE CPGH FGHI ELAK PVFH VGFL VKTM KVLR CVCF FCSK LLVD SNNP KIKD ILAK SKGQ PKKR LTHV YDLC KGKN ICEG GEEM DNKF GVEQ PEGD EDLT KEKG HGGC GRYQ PRIR RSGL ELYA EWKH VNED SQEK KILL SPER VHEI FKRI SDEE CFVL GMEP RYAR PEWM IVTV LPVP PLSV
Gene ID:	5430
Uniprot:	P24928
Cellular Location:	Nucleus
Calculated MW:	63kDa/217kDa
Observed MW:	250kDa

Synonyms:

POLR2A, POLR2, POLRA, RPB1, RPBh1, RPO2, RPOL2, RpIILS, hRPB220, hsRPB1, Pol II

Background:

This gene encodes the largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. The product of this gene contains a carboxy terminal domain composed of heptapeptide repeats that are essential for polymerase activity. These repeats contain serine and threonine residues that are phosphorylated in actively transcribing RNA polymerase. In addition, this subunit, in combination with several other polymerase subunits, forms the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA.

UniProt Protein Function:	POLR2A: DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relative to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. During transcription elongation, Pol II moves on the template as the transcript elongates. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Acts as a RNA- dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicate and transcriptase for the viral RNA circular genome. Component of the RNA polymerase II (Pol II) complex consisting of 12 subunits. The phosphorylated C-terminal domain interacts with FNBP3 and SYNCRIP. Interacts with SAFB/SAFB1. Interacts with CCNL1 and MYO1C. Interacts with CCNL2 and SFRS19. Component of a complex which is at least composed of HTATSF1/Tat-SF1, the P-TEFb complex components CDK9 and CCNT1, RNA polymerase II, SUPT5H, and NCL/nucleolin. Interacts with PAF1. Interacts (via C-terminus) with FTSJD2, CTDSP1 and SCAF8. Interacts via the phosphorylated C-terminal domain with WDR82 and with SETD1A and SETD1B only in the presence of WDR82. Interacts with ATF7IP. When phosphorylated at 'Ser-5', interacts with MEN1; the unphosphorylated form, or phosphorylated at 'Ser-2' does not interact. Interacts with DDX5. Belongs to the RNA polymerase beta' chain family.
UniProt Protein Details:	Protein type:Transferase; EC 2.7.7.6; Nucleotide Metabolism - purine; Transcription initiation complex; EC 2.7.7.48; Nucleotide Metabolism - pyrimidine Chromosomal Location of Human Ortholog: 17p13.1 Cellular Component: DNA-directed RNA polymerase II, core complex; nucleolus; nucleoplasm; nucleus Molecular Function:DNA binding; DNA-directed RNA polymerase activity; metal ion binding; protein binding; RNA-directed RNA polymerase activity; ubiquitin protein ligase binding Biological Process: DNA repair; gene expression; mRNA capping; nuclear mRNA splicing, via spliceosome; nucleotide-excision repair; positive regulation of RNA splicing; positive regulation of viral transcription; regulation of transcription, DNA-dependent; RNA elongation from RNA polymerase II promoter; RNA splicing; RNA-mediated gene silencing; somatic stem cell maintenance; transcription from RNA polymerase II promoter; transcription initiation from RNA polymerase II promoter; transcription termination; transcription-coupled nucleotide-excision repair; viral reproduction
NCBI Summary:	This gene encodes the largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. The product of this gene contains a carboxy terminal domain composed of heptapeptide repeats that are essential for polymerase activity. These repeats contain serine and threonine residues that are phosphorylated in actively transcribing RNA polymerase. In addition, this subunit, in combination with several other polymerase subunits, forms the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA. [provided by RefSeq, Jul 2008]
UniProt Code:	P24928
NCBI GenInfo Identifier:	281185484
NCBI Gene ID:	5430
NCBI Accession:	P24928.2
UniProt Secondary Accession:	P24928,Q6NX41, A6NN93, B9EH88,
UniProt Related Accession:	P24928
Molecular Weight:
NCBI Full Name:	DNA-directed RNA polymerase II subunit RPB1
NCBI Synonym Full Names:	polymerase (RNA) II subunit A
NCBI Official Symbol:	POLR2AÂ Â
NCBI Official Synonym Symbols:	RPB1; RPO2; POLR2; POLRA; RPBh1; RPOL2; RpIILS; hsRPB1; hRPB220Â Â
NCBI Protein Information:	DNA-directed RNA polymerase II subunit RPB1
UniProt Protein Name:	DNA-directed RNA polymerase II subunit RPB1
UniProt Synonym Protein Names:	DNA-directed RNA polymerase II subunit A; DNA-directed RNA polymerase III largest subunit; RNA-directed RNA polymerase II subunit RPB1 (EC:2.7.7.48)
UniProt Gene Name:	POLR2AÂ Â
UniProt Entry Name:	RPB1_HUMAN

	Western blot analysis of extracts of various cell lines, using POLR2A antibody at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 30s.
	Immunohistochemistry of paraffin-embedded human esophageal using POLR2A Rabbit pAb at dilution of 1:50 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with IHC staining protocol.
	Immunohistochemistry of paraffin-embedded mouse spleen using POLR2A Rabbit pAb at dilution of 1:50 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with IHC staining protocol.
	Immunohistochemistry of paraffin-embedded rat lung using POLR2A Rabbit pAb at dilution of 1:50 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with IHC staining protocol.
	Immunoprecipitation analysis of 200ug extracts of HeLa cells using 3ug POLR2A antibody at a dilition of 1:1000.
	Chromatin immunoprecipitation analysis of extracts of HeLa cells, using POLR2A antibody and rabbit IgG. The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.

Antibodies	ELISA Kits
Anti-POLR2A Antibody (CAB2107)	POLR2A Colorimetric Cell-Based ELISA Kit
Anti-Phospho-POLR2A-S2 pAb Antibody (CABP0749)	POLR2A (Phospho-Ser1619) Colorimetric Cell-Based ELISA Kit
Anti-Phospho-POLR2A-S5 pAb Antibody (CABP0828)
Anti-Phospho-POLR2A-S2 Antibody (CABP0996)
Anti-Phospho-POLR2A-S5 Antibody (CABP0997)

Secondary Antibody

Anti-HRP Goat Anti-Rabbit IgG (H+L) Antibody (CABS014)