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ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set
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REFERENCES
WNT10B/β-catenin signalling induces HMGA2 and proliferation in metastatic triple-negative breast cancer.
Wend, Peter, et al. (2013) EMBO Mol Med, 5: 264-79 (2013)
Transcriptional regulation of Nox4 by histone deacetylases in human endothelial cells.
Daniel Siuda,Ulrich Zechner,Nady El Hajj,Dirk Prawitt,David Langer,Ning Xia,Sven Horke,Andrea Pautz,Hartmut Kleinert,Ulrich Förstermann,Huige Li (2012) Basic research in cardiology.107
The telomeric transcriptome of Schizosaccharomyces pombe.
Amadou Bah,Harry Wischnewski,Vadim Shchepachev,Claus M Azzalin (2012) Nucleic acids research.40
Species Reactivity Key Applications Host Format Antibody Type
H, M, R, Yeast (S. cerevisiae) WB, IHC, ICC, ChIP Mouse Ascites Monoclonal Antibody
Description:
ChIPAb+ RNA Pol II - ChIP Validated Antibody and Primer Set
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Trade Name:
Upstate (Millipore)
Product Overview:
All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ RNA Polymerase II (RNA Pol II) set includes the Anti-RNA Pol II, clone 8WG16 antibody, a negative control monoclonal antibody, and qPCR primers which amplify a 166 bp region within the promoter of the human GAPDH gene. The RNA Pol II clone 8WG16 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of RNA polymerase II-associated chromatin.
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Specificity:
Recognizes RNA polymerase II, Mr ~220 kDa.
Molecular Weight:
~220 kDa
Epitope:
C-terminus of large subunit of Pol II
Immunogen:
RNA Polymerase II purified from wheat germ extract.
Clone:
8WG16
Isotype:
IgG2a
Species Reactivity:
Human
Mouse
Rat
Yeast (S. cerevisiae)
Species Reactivity Note:
Human, mouse, rat, yeast. Not tested in other species, but broad species reactivity is expected based on conservation of the C-terminal region of RNA pol II.
Application Notes:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 μl of either Negative Ascites or anti-RNA Pol II clone 8WG16 antibody and the Magna ChIP G Kit (Part No. 17-611).
Successful immunoprecipitation of RNA Pol II associated DNA fragments was verified by qPCR using Control Primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
1:500-1:2000 dilution of a previous lot detected RNA Polymerase II in HeLa nuclear extract.
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Control:
Includes negative ascites and control primers specific for human GAPDH promoter.
Quality Assurance:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µl of either a Ne |