NB200-598

antibody from Novus Biologicals

Targeting: POLR2A POLR2, POLRA, RPB1

Western blot (Supportive data in Antibodypedia)

ELISA (Data presented on provider website)

Immunocytochemistry (Data presented on provider website)

Immunoprecipitation (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Data presented on provider website)

Antibody Data

Product number

NB200-598

Provider

Novus Biologicals

Proper citation

Novus Cat#NB200-598, RRID:AB_2167465

Product name

Mouse Monoclonal RNA Polymerase II/POLR2A Antibody

Antibody type

Monoclonal

Description

Protein G purified. 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

Reactivity

Human, Mouse, Simian, Yeast

Host

Mouse

Isotype

IgG

Vial size

0.1 ml

Concentration

1 mg/ml

Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

References

Submitted references

Cyclic Multiplexed-Immunofluorescence (cmIF), a Highly Multiplexed Method for Single-Cell Analysis.

Eng J, Thibault G, Luoh SW, Gray JW, Chang YH, Chin K
Methods in molecular biology (Clifton, N.J.) 2020;2055:521-562

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Sen1 Is Recruited to Replication Forks via Ctf4 and Mrc1 and Promotes Genome Stability.

Appanah R, Lones EC, Aiello U, Libri D, De Piccoli G
Cell reports 2020 Feb 18;30(7):2094-2105.e9

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Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes.

Lin JR, Izar B, Wang S, Yapp C, Mei S, Shah PM, Santagata S, Sorger PK
eLife 2018 Jul 11;7

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PRDX2 and PRDX4 are negative regulators of hypoxia-inducible factors under conditions of prolonged hypoxia.

Luo W, Chen I, Chen Y, Alkam D, Wang Y, Semenza GL
Oncotarget 2016 Feb 9;7(6):6379-97

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Spt6 prevents transcription-coupled loss of posttranslationally modified histone H3.

Kato H, Okazaki K, Iida T, Nakayama J, Murakami Y, Urano T
Scientific reports 2013;3:2186

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Peripheral subnuclear positioning suppresses Tcrb recombination and segregates Tcrb alleles from RAG2.

Chan EA, Teng G, Corbett E, Choudhury KR, Bassing CH, Schatz DG, Krangel MS
Proceedings of the National Academy of Sciences of the United States of America 2013 Nov 26;110(48):E4628-37

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Plasma membrane associated transcription of cytoplasmic DNA.

Cheng J, Torkamani A, Peng Y, Jones TM, Lerner RA
Proceedings of the National Academy of Sciences of the United States of America 2012 Jul 3;109(27):10827-31

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Proteasome regulation of dynamic transcription factor occupancy on the GnRH-stimulated luteinizing hormone beta-subunit promoter.

Walsh HE, Shupnik MA
Molecular endocrinology (Baltimore, Md.) 2009 Feb;23(2):237-50

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DNA damage-induced Def1-RNA polymerase II interaction and Def1 requirement for polymerase ubiquitylation in vitro.

Reid J, Svejstrup JQ
The Journal of biological chemistry 2004 Jul 16;279(29):29875-8

Western blot

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

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.

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

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 and probed with 2 ug/mL NB200-598. RNA Pol II reactivity (arrowhead) is observed in HeLa, MCF and 3T3 cells.

Flow cytometry

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

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 NBP2-27287 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).

Supportive validation

Submitted by

Novus Biologicals (provider)

Main image

Experimental details

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 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes. Both antibodies were conjugated to phycoerythrin.