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  • ZMS1031 - Anti-RNA polymerase II Antibody, clone CTD4H8 ZooMAb® Mouse Monoclonal

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ZMS1031 Sigma-Aldrich

Anti-RNA polymerase II Antibody, clone CTD4H8 ZooMAb® Mouse Monoclonal

New Green Alternative Antibody Enhanced Validation

recombinant, expressed in HEK 293 cells

Synonym: 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, RNA polymerase II subunit B1, RNA-directed RNA polymerase II subunit RPB1

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Properties

Related Categories 12 Principles Aligned Products, Alphabetical Index, Antibodies, Antibodies (Greener), Chemical Synthesis,
product line   ZooMAb® learn more
conjugate   unconjugated
clone   CTD4H8, monoclonal
  recombinant monoclonal
biological source   mouse
application(s)   flow cytometry: suitable
  immunocytochemistry: suitable
  immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
  western blot: suitable
species reactivity   human
species reactivity (predicted by homology)   mouse, yeast, rat
mol wt   calculated mol wt 217.18 kDa
  observed mol wt ~300 kDa
enhanced validation   recombinant expression
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form   lyophilized
shipped in   ambient
storage temp.   2-8°C
packaging   antibody small pack of 25 μL
antibody form   purified antibody
isotype   IgG1
Quality Level   200
recombinant   expressed in HEK 293 cells
antibody product type   primary antibodies
greener alternative product characteristics   Waste Prevention
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greener alternative category   Aligned

Description

General description



ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies.

Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them.

Learn more about ZooMAb here.

Specificity

Clone CTD4H8 is a ZooMAb mouse recombinant monoclonal antibody that specifically detects RNA polymerase II. It targets an epitope within the C-terminal region.

Immunogen

Peptide containing 10 repeats of YSPT[pS]PS corresponding to the C-terminal region of human RNA polymerase II.

Application

Anti-RNA polymerase II, clone CTD4H8 ZooMAb, Cat. No. ZMS1031, is a recombinant Mouse monoclonal antibody that detects RNA polymerase II and is tested in Flow Cytometry, Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting.

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected RNA polymerase II in HeLa and NIH3T3 cell lysate.

Flow Cytometry Analysis: 0.1 μg from a representative lot detected RNA polymerase II in one million HUVEC.

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected RNA polymerase II in human cerebral cortex and human testis tissue sections.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected RNA polymerase II in HeLa, A431, HUVEC, and NIH 3T3 cells.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Target description

DNA-directed RNA polymerase II subunit RPB1 (UniProt: P24928; also known as EC:2.7.7.6; RNA polymerase II subunit B1, DNA-directed RNA polymerase II subunit A, DNA-directed RNA polymerase III largest subunit, RNA-directed RNA polymerase II subunit RPB1, POLR2A) is encoded by the POLR2A (also known as POLR2) gene (Gene ID: 5430) in human. DNA-directed RNA polymerase II subunit RPB1 is the catalytic component of RNA polymerase II that catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. It has multiple Zinc and magnesium ion binding sites. The binding of ribonucleoside triphosphate to the RNA polymerase II transcribing complex 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 nucleotide 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. POLR2A is localized to cytoplasm in its hypo-phosphorylated form and the active hyper-phosphorylated form is located mainly in the nucleus. The tandem heptapeptide repeats (Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7) in its C-terminal domain (CTD) can be highly phosphorylated and regulates transcription coupled processes. Phosphorylation is shown to occur mainly at serine 2 and serine 5 residues of the heptapeptide repeat and is mediated by CDK7 and CDK9. Phosphorylation can also take place at serine 7 of the heptapeptide repeat, which is required for efficient transcription of snRNA genes and processing of the transcripts. Its C-terminal domain (aa 1593-1960) serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. The K7-residues in non-consensus repeats of human RNAPII are modified by acetylation, or mono-, di-, and trimethylation. Acetylation of K7, and di- and tri-methylation of K7 are found exclusively associated with phosphorylated CTD peptides. Monomethylated K7 is found in non-phosphorylated CTD. Clone 3G21 targets both phosphorylated and non-phosphorylated YSPTSPS repeats in the C-terminal region. Q403K or L438H mutations in POLR2A are reported to hijack RNA polymerase II and drive neoplasia and POLR2A mutant tumors display dysregulation of key meningeal identity genes, including WNT6 and ZIC1/ZIC4. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Voss, K., et al. (2015). Transcription 6(5); 91-101; Clark, VE., et al. (2016). Nat. Gen. 48(10); 1253 1259).

Physical form

Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.

Reconstitution

30 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Disclaimer

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.

Legal Information

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Safety & Documentation

Safety Information

WGK Germany 
WGK 3
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

Antibody Basics

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