RNA Polymerase II/POLR2A Antibody (4H8)

Images

 
Western Blot: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - Whole cell protein from HeLa (lane 1), MCF7 (lane 2), NIH-3T3 (lane 3) and Cos7 (lane 4) were separated by SDS-PAGE on a 6% gel, transferred to PVDF ...read more
Immunocytochemistry/ Immunofluorescence: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - RNA Polymerase II/POLR2A antibody was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were ...read more
Flow Cytometry: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - An intracellular stain was performed on HeLa cells with RNA Polymerase II/POLR2A (4H8) antibody NB200-598PE (blue) and a matched isotype control ...read more
Western Blot: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - Analysis of MCF7 cell lysata (A) using RNA polymerase II antibody at 2 ug/mL.
Flow (Intracellular): RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - An intracellular stain was performed on Daudi cells with RNA Polymerase II/POLR2A (4H8) antibody NB200-598 (blue) and a matched isotype control ...read more

Product Details

Summary
Reactivity Hu, Mu, Ye, Pm(-), Dr, Ha, PlSpecies Glossary
Applications WB, ChIP, ELISA, Flow, Flow-IC, ICC/IF, IP, CyTOF-ready
Clone
4H8
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1 mg/ml

RNA Polymerase II/POLR2A Antibody (4H8) Summary

Immunogen
10 repeats of synthetic peptide YSPTSPS using chemically synthesized phospho-Ser 5 YSPTSpPS (Human). [UniProt# P24928]
Specificity
Please note that this antibody detects both unphosphorylated and phosphorylated forms of the protein. When using this antibody in western blot and IP, it recognizes both the II0 and IIA form of RPB1 (ie the largest RNAPII subunit). These different forms of RPB1 (as seen in gels) have traditionally been loosely defined as 'hyper-' and 'hypo-phosphorylated', respectively. Here, however, it is important to remember that RPB1 contains no less than 52 repeats of a heptapeptide (TSPTSPS) that can be phosphorylated at different positions (Tyr1, Ser2, Thr4, Ser5 and Ser7; this antibody was raised against a synthetic peptide with 10 of these repeats, mostly but probably not entirely, chemically phosphorylated at Ser5). So, when we say that it recognizes both forms (II0/hyper-phos and IIA/hypo-phos), it might really just mean that the antibody recognizes a few phosphorylated Ser5s among 52 otherwise mostly un-phosphorylated repeats. Alternatively, it might conceivably (though less likely) recognize UN-phosphorylated repeats as there might conceivably be a few of those even in the II0/hyper-phos RPB1 form.'
Predicted Species
Drosophila (100%), Hamster (100%), Plant (100%). Backed by our 100% Guarantee.
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
POLR2A
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1 - 2 ug/mL
  • Chromatin Immunoprecipitation 1:10 - 1:500
  • ELISA 1:100 - 1:2000
  • Flow Cytometry 1 ug per million cells
  • Immunocytochemistry/Immunofluorescence 1:100 - 1:1000
  • Immunoprecipitation 1:10 - 1:500
  • CyTOF-ready
  • Flow (Intracellular)
Application Notes
In WB a band can be seen at approximately 217 kDa. Optimal dilutions/concentrations should be determined by the end user. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Theoretical MW
217 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Control
NIH 3T3 Lysate (NB800-PC8)
Reviewed Applications
Read 1 Review rated 5
using
NB200-598 in the following applications:

Publications
Read Publications using
NB200-598 in the following applications:

  • 2 publications
  • 3 publications
  • IP
    1 publication
  • WB
    3 publications

Reactivity Notes

Does not cross react with monkey.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
No Preservative
Concentration
1 mg/ml
Purity
Protein G purified

Alternate Names for RNA Polymerase II/POLR2A Antibody (4H8)

  • DNA-directed RNA polymerase II largest subunit, RNA polymerase II 220 kdsubunit
  • DNA-directed RNA polymerase II subunit A
  • DNA-directed RNA polymerase II subunit RPB1
  • DNA-directed RNA polymerase III largest subunit
  • EC 2.7.7.48
  • EC 2.7.7.6
  • hRPB220
  • hsRPB1
  • MGC75453
  • POLR2
  • POLR2A
  • POLRA
  • polymerase (RNA) II (DNA directed) polypeptide A (220kD)
  • polymerase (RNA) II (DNA directed) polypeptide A, 220kDa
  • RNA Pol 2
  • RNA Polymerase 2
  • RNA polymerase II subunit B1
  • RNA Polymerase II
  • RNA-directed RNA polymerase II subunit RPB1
  • RNAPII
  • RPB1
  • RPBh1
  • RpIILS
  • RPO2
  • RPOL2

Background

RNA polymerase II (RNAPII) is a 12-subunit enzyme which synthesizes all mRNAs as well as several non-coding RNAs in eukaryotic cells thereby playing a central role in cellular functions. RNAPII is a DNA-dependent RNA polymerase enzyme which catalyzes the transcription of DNA into RNA employing four ribonucleoside triphosphates as substrates. The activity of RNAPII transcription machinery is regulated by DNA-binding transcriptional regulators in a gene/cell type-specific manner, whereas general transcription factors act as RNAPII accessory proteins required for the transcription of most class II genes, and the co-activators/co-repressors serve as bridges between DNA-bound factors and the RNAPII machinery, some affecting the organization and/or chemical modification of the chromatin template of RNAPII. RNA polymerase II subunit B1 (POLR2A) is the largest and the catalytic component of RNAPII which forms the polymerase active center together with the second largest subunit and is the central component of the basal RNAPII transcription machinery. It is composed of mobile elements that move relative to each other and POLR2A is the 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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NB200-598
Species: Hu, Mu, Ye, Pm(-), Dr, Ha, Pl
Applications: WB, ChIP, ELISA, Flow, Flow-IC, ICC/IF, IP, CyTOF-ready

Publications for RNA Polymerase II/POLR2A Antibody (NB200-598)(7)

We have publications tested in 3 confirmed species: Human, Mouse, Yeast.

We have publications tested in 4 applications: ChIP, ICC/IF, IP, WB.


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ChIP
(2)
ICC/IF
(3)
IP
(1)
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(3)
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Human
(3)
Mouse
(1)
Yeast
(2)
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Showing Publications 1 - 7 of 7.
Publications using NB200-598 Applications Species
Lin JR, Izar B, Wang S et al. Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes Elife Jul 11 2018 [PMID: 29993362] (ICC/IF, Human) ICC/IF Human
Luo W, Chen I, Chen Y et al. PRDX2 and PRDX4 are negative regulators of hypoxia-inducible factors under conditions of prolonged hypoxia. Oncotarget 2016 Feb 9 [PMID: 26837221] (WB) WB
Chan EA, Teng G, Corbett E et al. Peripheral subnuclear positioning suppresses Tcrb recombination and segregates Tcrb alleles from RAG2. Proc Natl Acad Sci U S A. 2013 Nov 26 [PMID: 24218622] (ICC/IF, Mouse) ICC/IF Mouse
Kato H, Okazaki K, Iida T et al. Spt6 prevents transcription-coupled loss of posttranslationally modified histone H3. Sci Rep 2013 Jul 15 [PMID: 23851719] (ChIP, Yeast) ChIP Yeast
Cheng J, Torkamani A, Peng Y et al. Plasma membrane associated transcription of cytoplasmic DNA Proc Natl Acad Sci U S A 2012 Jun 18 [PMID: 22711823] (WB, ICC/IF, Human) WB, ICC/IF Human
Walsh HE Shupnik MA. Proteasome regulation of dynamic transcription factor occupancy on the GnRH-stimulated luteinizing hormone beta-subunit promoter. Mol Endocrinol;23(2):237-50. 2009 Feb. [PMID: 19095772] (ChIP, Human) ChIP Human
Reid J, Svejstrup JQ. DNA damage-induced Def1-RNA polymerase II interaction and Def1 requirement for polymerase ubiquitylation in vitro. J Biol Chem;279(29):29875-8. 2004 Jul 16. [PMID: 15166235] (WB, IP, Yeast) WB, IP Yeast

Review for RNA Polymerase II/POLR2A Antibody (NB200-598) (1) 51

Average Rating: 5
(Based on 1 review)
We have 1 review tested in 1 species: Human.

Reviews using NB200-598:
Filter by Applications
WB
(1)
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Human
(1)
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Images Ratings Applications Species Date Details
Western Blot RNA Polymerase II/POLR2A NB200-598
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reviewed by:
Anony- mous
WB Human 05/13/2014
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Summary

ApplicationWestern Blot
SpeciesHuman
Special ApplicationsSubstrate of fourteen repeats of YSPTSPS fused to GST for purification (~50kD) from bacteria. We tested the ability of four purified yeast kinases to phosphorylate the Ser5 residue of our CTD substrate in vitro. Western Blot is probed with 4H8 Alexa 647 antibody.

Product General Protocols

View specific protocols for RNA Polymerase II/POLR2A Antibody (NB200-598): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ChIP Webinar
ChIP Video Protocol
ICC/IF Video Protocol

FAQs for RNA Polymerase II/POLR2A Antibody (NB200-598). (Showing 1 - 1 of 1 FAQs).

  1. The immunogen info mentions 10 repeats of synthetic peptide YSPTSPS. What does this mean?
    • It is correct that the immunogen is literally 10 adjacent repeats of the sequence YSPTSPS, where the Ser 5 of each repeat is phosphorylated (the 'p Ser5' in the product name referring to serine 5 of this repeat rather than of the whole target protein). The datasheet for NB200-598 refers to UniProt #P24928. If you look at this sequence, you will see that it does contain these repeats of YSPTSPS; I ran a sequence comparison of this protein and 10x YSPTSPS and it aligned perfectly. For further information, the 'Background' section at the bottom of the datasheet for NB200-598 says, 'Recruitment of RNA processing factors to the elongation complex is coordinated by the phosphorylation of Serine-2 and Serine-5 residues within the C-terminal repeat (YSPTSPS) domain of the largest subunit of RNAPII'; and the 'Specificity' section near the top says, 'This antibody recognizes the C-terminal repeat of the largest subunit of RNA polymerase II. It recognizes both unphosphorylated and phosphorylated forms'.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB200-598AF405
Alexa Fluor 488 Labeled NB200-598AF488
Alexa Fluor 647 Labeled NB200-598AF647
Alexa Fluor 700 Labeled NB200-598AF700
Allophycocyanin Labeled NB200-598APC
Biotin Labeled NB200-598B
DyLight 350 Labeled NB200-598UV
DyLight 405 Labeled NB200-598V
DyLight 488 Labeled NB200-598G
DyLight 550 Labeled NB200-598R
DyLight 650 Labeled NB200-598C
DyLight 680 Labeled NB200-598FR
DyLight 755 Labeled NB200-598IR
FITC Labeled NB200-598F
HRP Labeled NB200-598H
PE Labeled NB200-598PE

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Recent Reviews

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Anony- mous
05/13/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol POLR2A
Entrez
Uniprot