Anti-RNA polymerase II subunit B1, clone 4F8 (rat monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
M, H	WB, ELISA, ChIP	R	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

M, H

WB, ELISA, ChIP

Purified

Monoclonal Antibody

Description
Catalogue Number	04-1569
Description	Anti-RNA polymerase II subunit B1 Antibody, clone 4F8
Alternate Names	DNA-directed RNA polymerase II A DNA-directed RNA polymerase II largest subunit, RNA polymerase II 220 kd subunit DNA-directed RNA polymerase II subunit A DNA-directed RNA polymerase III largest subunit RNA polymerase II subunit B1 RNA-directed RNA polymerase II subunit RPB1 polymerase (RNA) II (DNA directed) polypeptide A (220kD) polymerase (RNA) II (DNA directed) polypeptide A, 220kDa
Background Information	RNA polymerase II subunit B1 (RPB1) is the largest subunit of the RNA polymerase II complex. As a holoenzyme RNA polymerase II catalyzes transcription of eukaryotic DNA into RNA using the four ribonucleoside triphosphates as substrates. The RBP1 subunit, in combination with other polymerase subunits, forms a large central cleft that maintains contact between the active site of the enzyme, the DNA template, and the nascent RNA transcript. This subunit also contains a carboxy terminal domain (CTD) consisting of tandem heptapeptide repeats. In actively transcribing RNA polymerase ‘Ser-2’ and ‘Ser-5’ of the heptapeptide repeat are phosphorylated. Phosphorylation activates the RNA polymerase II beta subunit, allowing it to serve as an assembly platform for additional subunits that modulate initiation, elongation, termination and mRNA processing.

Description

Catalogue Number

04-1569

Description

Anti-RNA polymerase II subunit B1 Antibody, clone 4F8

Alternate Names

DNA-directed RNA polymerase II A
DNA-directed RNA polymerase II largest subunit, RNA polymerase II 220 kd subunit
DNA-directed RNA polymerase II subunit A
DNA-directed RNA polymerase III largest subunit
RNA polymerase II subunit B1
RNA-directed RNA polymerase II subunit RPB1
polymerase (RNA) II (DNA directed) polypeptide A (220kD)
polymerase (RNA) II (DNA directed) polypeptide A, 220kDa

Background Information

RNA polymerase II subunit B1 (RPB1) is the largest subunit of the RNA polymerase II complex. As a holoenzyme RNA polymerase II catalyzes transcription of eukaryotic DNA into RNA using the four ribonucleoside triphosphates as substrates. The RBP1 subunit, in combination with other polymerase subunits, forms a large central cleft that maintains contact between the active site of the enzyme, the DNA template, and the nascent RNA transcript. This subunit also contains a carboxy terminal domain (CTD) consisting of tandem heptapeptide repeats. In actively transcribing RNA polymerase ‘Ser-2’ and ‘Ser-5’ of the heptapeptide repeat are phosphorylated. Phosphorylation activates the RNA polymerase II beta subunit, allowing it to serve as an assembly platform for additional subunits that modulate initiation, elongation, termination and mRNA processing.

Product Information
Format	Purified
Control	NIH/3T3 cell lysates
Presentation	Purified rat monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4, 150 mM NaCl) with 0.05% sodium azide.

Product Information

Format

Purified

Control

NIH/3T3 cell lysates

Presentation

Purified rat monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4, 150 mM NaCl) with 0.05% sodium azide.

Applications
Application	Anti-RNA polymerase II subunit B1 Antibody, clone 4F8 is a Rat monoclonal antibody for detection of RNA polymerase II subunit B1 also known as DNA-directed RNA polymerase II A & has been validated in WB, ELISA.
Key Applications	Western Blotting ELISA Chromatin Immunoprecipitation (ChIP)
Application Notes	Chromatin Immunoprecipitation Analysis: A representative lot was used by independent laboraotry in ChIP. (Chapman, R., et al. (2007). Science. 318(5857):1780 -1782.)

Applications

Application

Anti-RNA polymerase II subunit B1 Antibody, clone 4F8 is a Rat monoclonal antibody for detection of RNA polymerase II subunit B1 also known as DNA-directed RNA polymerase II A & has been validated in WB, ELISA.

Key Applications

Western Blotting
ELISA
Chromatin Immunoprecipitation (ChIP)

Application Notes

Chromatin Immunoprecipitation Analysis: A representative lot was used by independent laboraotry in ChIP. (Chapman, R., et al. (2007). Science. 318(5857):1780 -1782.)

Biological Information
Immunogen	Ovalbumin-conjugated linear peptide corresponding to human RNA polymerase II subunit B1.
Epitope	Unknown
Clone	4F8
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Rat
Specificity	This antibody recognizes RNA polymerase II subunit B1.
Isotype	IgG1κ
Species Reactivity	Mouse Human
Species Reactivity Note	Demonstrated to react with mouse. Predicted to react with human based on 100% sequence homology.
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_000928
Entrez Gene 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].
Gene Symbol	POLR2A POLR2 POLRA RPB1 RPBh1 RPO2 RPOL2 RpIILS hRPB220 hsRPB1
Purification Method	Protein G Purified
UniProt Number	P24928
UniProt Summary	FUNCTION: 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. CATA;YTIC ACTIVITY: Nucleoside triphosphate + RNA(n) = diphosphate + RNA(n+1). SUBUNIT STRUCTURE: 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 By similarity. 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. SUBCELLULAR LOCATION: Nucleus PTM: The tandem 7 residues repeats in the C-terminal domain (CTD) can be highly phosphorylated. The phosphorylation activates Pol II. Phosphorylation occurs mainly at residues 'Ser-2' and 'Ser-5' of the heptapepdtide repeat. The phosphorylation state is believed to result from the balanced action of site-specific CTD kinases and phosphataes, and a "CTD code" that specifies the position of Pol II within the transcription cycle has been proposed. MISCELLANEOUS: The binding of ribonucleoside triphosphate to the RNA polymerase II transcribing complex probably involves a two-step mechanism. The initial binding seems to occur at the entry (E) site and involves a magnesium ion temporarily coordinated by three conserved aspartate residues of the two largest RNA Pol II subunits. The ribonucleoside triphosphate is transferred by a rotation to the nucelotide addition (A) site for pairing with the template DNA. The catalytic A site involves three conserved aspartate residues of the RNA Pol II largest subunit which permanently coordinate a second magnesium ion. SEQUENCE SIMILARITIES: Belongs to the RNA polymerase beta' chain family.
Molecular Weight	~ 220 kDa

Biological Information

Immunogen

Ovalbumin-conjugated linear peptide corresponding to human RNA polymerase II subunit B1.

Epitope

Unknown

Clone

4F8

Concentration

Please refer to the Certificate of Analysis for the lot-specific concentration.

Host

Rat

Specificity

This antibody recognizes RNA polymerase II subunit B1.

Isotype

IgG1κ

Species Reactivity

Mouse
Human

Species Reactivity Note

Demonstrated to react with mouse. Predicted to react with human based on 100% sequence homology.

Antibody Type

Monoclonal Antibody

Entrez Gene Number

NP_000928

Entrez Gene 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].

Gene Symbol

POLR2A
POLR2
POLRA
RPB1
RPBh1
RPO2
RPOL2
RpIILS
hRPB220
hsRPB1

Purification Method

Protein G Purified

UniProt Number

P24928

UniProt Summary

FUNCTION: 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.

CATA;YTIC ACTIVITY: Nucleoside triphosphate + RNA(n) = diphosphate + RNA(n+1).

SUBUNIT STRUCTURE: 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 By similarity. 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.

SUBCELLULAR LOCATION: Nucleus

PTM: The tandem 7 residues repeats in the C-terminal domain (CTD) can be highly phosphorylated. The phosphorylation activates Pol II. Phosphorylation occurs mainly at residues 'Ser-2' and 'Ser-5' of the heptapepdtide repeat. The phosphorylation state is believed to result from the balanced action of site-specific CTD kinases and phosphataes, and a "CTD code" that specifies the position of Pol II within the transcription cycle has been proposed.

MISCELLANEOUS: The binding of ribonucleoside triphosphate to the RNA polymerase II transcribing complex probably involves a two-step mechanism. The initial binding seems to occur at the entry (E) site and involves a magnesium ion temporarily coordinated by three conserved aspartate residues of the two largest RNA Pol II subunits. The ribonucleoside triphosphate is transferred by a rotation to the nucelotide addition (A) site for pairing with the template DNA. The catalytic A site involves three conserved aspartate residues of the RNA Pol II largest subunit which permanently coordinate a second magnesium ion.

SEQUENCE SIMILARITIES: Belongs to the RNA polymerase beta' chain family.

Molecular Weight

~ 220 kDa

Product Usage Statements
Quality Assurance	Evaluated by Western Blot in NIH/3T3 cell lysates. Western Blot Analysis: 1 µg/ml of this antibody detected RNA polymerase II subunit B1 on 10 µg of NIH/3T3 cell lysates.
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Quality Assurance

Evaluated by Western Blot in NIH/3T3 cell lysates.

Western Blot Analysis: 1 µg/ml of this antibody detected RNA polymerase II subunit B1 on 10 µg of NIH/3T3 cell lysates.

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Stable for 1 year at 2-8°C from date of receipt.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at 2-8°C from date of receipt.

Packaging Information
Material Size	100 µg

Packaging Information

Material Size

100 µg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Anti-RNA polymerase II subunit B1, clone 4F8 (rat monoclonal)

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