Chromatin Immunoprecipitation (ChIP)
(RNA pol II CTD K7me1 antibody (pAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT High Sensitivity Kit with 30 ug of chromatin from Raji cells and 10 ug of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 10.9M sequence tags were mapped to identify RNA pol II binding sites across a region of chromosome 12.)
Rat anti-Human RNA pol II CTD monomethyl Lys7 Monoclonal Antibody | anti-Lys7 antibody
RNA pol II CTD monomethyl Lys7 antibody (mAb)(Clone 1F5)
ChIP-Seq: 10ug per ChIP
IP: 5ug per IP
WB: 0.5-2ug/ml
Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt/Sonication Protocol when preparing nuclear extracts for Western blot.
Chromatin Immunoprecipitation (ChIP)
(RNA pol II CTD K7me1 antibody (pAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT High Sensitivity Kit with 30 ug of chromatin from Raji cells and 10 ug of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 10.9M sequence tags were mapped to identify RNA pol II binding sites across a region of chromosome 12.)
Chromatin Immunoprecipitation (ChIP)
(RNA pol II CTD K7me1 antibody (pAb) tested by ChIP-Seq. ChIP was performed using the ChIP-IT High Sensitivity Kit with 30 ug of chromatin from Raji cells and 10 ug of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 10.9M sequence tags were mapped to identify RNA pol II binding sites across a region of chromosome 12.)
Western Blot (WB)
(RNA pol II CTD monomethyl Lys7 antibody tested by Western blot. 20 ug of K562 nuclear extract* was run on SDS-PAGE and probed with antibody at 0.5 ug/ml.)
Western Blot (WB)
(RNA pol II CTD monomethyl Lys7 antibody tested by Western blot. 20 ug of K562 nuclear extract* was run on SDS-PAGE and probed with antibody at 0.5 ug/ml.)
Immunoprecipitation (IP)
(RNA pol II CTD monomethyl Lys7 antibody tested by Immunoprecipitation. 5 ug of antibody was used to immunoprecipitate RNA pol II CTD monomethyl Lys7 from 300 ug of K562 nuclear extract, which was then run on SDS-PAGE and probed with the same antibody at 3 ug/ml.)
Immunoprecipitation (IP)
(RNA pol II CTD monomethyl Lys7 antibody tested by Immunoprecipitation. 5 ug of antibody was used to immunoprecipitate RNA pol II CTD monomethyl Lys7 from 300 ug of K562 nuclear extract, which was then run on SDS-PAGE and probed with the same antibody at 3 ug/ml.)
Testing Data
(RNA pol II CTD monomethyl Lys7 antibody tested for specificity Biotinylated peptides containing various RNA Pol II CTD Lysine 7 modifications were captured onto wells of a Streptavidin-coated plate and then incubated with RNA Pol II CTD K7me1 antibody at 1 ug/mL. Peptide-bound antibody was detected using anti-rat IgG HRP followed by addition of peroxidase substrate and read at A405.)
Testing Data
(RNA pol II CTD monomethyl Lys7 antibody tested for specificity Biotinylated peptides containing various RNA Pol II CTD Lysine 7 modifications were captured onto wells of a Streptavidin-coated plate and then incubated with RNA Pol II CTD K7me1 antibody at 1 ug/mL. Peptide-bound antibody was detected using anti-rat IgG HRP followed by addition of peroxidase substrate and read at A405.)
Background: Genome-wide profiling revealed high levels of K7me1/2 marks at the transcriptional start site of genes for sense and antisense transcribing RNAPII. The new K7 modifications further expand the mammalian CTD code to allow regulation of differential gene expression. In analogy, it appears likely that the various modifications of K7 residues identified in this analysis may similarly contribute to fine tuning of gene expressing by RNAPII, and that these modifications are placed in K7 residues of CTD dependent on cell signaling and epigenetic programming of cells.
NCBI and Uniprot Product Information
Uniprot Description
POLR2A: 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. 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. 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, CTDSP1 and SCAF8. Interacts via the phosphorylated C-terminal domain with WDR82 and with SETD1A and SETD1B only in the presence of WDR82. Interacts with ATF7IP. When phosphorylated at 'Ser-5', interacts with MEN1; the unphosphorylated form, or phosphorylated at 'Ser-2' does not interact. Interacts with DDX5. Belongs to the RNA polymerase beta' chain family.
Protein type: Nucleotide Metabolism - purine; Nucleotide Metabolism - pyrimidine; Transferase; EC 2.7.7.6; Transcription initiation complex; EC 2.7.7.48
Chromosomal Location of Human Ortholog: 17p13.1
Cellular Component: nucleoplasm; nucleolus; DNA-directed RNA polymerase II, core complex; nucleus
Molecular Function: protein binding; RNA-directed RNA polymerase activity; DNA binding; DNA-directed RNA polymerase activity; ubiquitin protein ligase binding; metal ion binding
Biological Process: transcription from RNA polymerase II promoter; transcription initiation from RNA polymerase II promoter; viral reproduction; somatic stem cell maintenance; positive regulation of viral transcription; RNA splicing; DNA repair; positive regulation of RNA splicing; nuclear mRNA splicing, via spliceosome; mRNA capping; regulation of transcription, DNA-dependent; nucleotide-excision repair; transcription-coupled nucleotide-excision repair; RNA elongation from RNA polymerase II promoter; gene expression
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Product Notes
The Lys7 polr2a (Catalog #AAA389059) is an Antibody produced from Rat and is intended for research purposes only. The product is available for immediate purchase. The RNA pol II CTD monomethyl Lys7 antibody (mAb)(Clone 1F5) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's RNA pol II CTD monomethyl Lys7 can be used in a range of immunoassay formats including, but not limited to, Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation-Seq (ChIP-Seq), Immunoprecipitation (IP), Western Blot (WB). Applications Validated: ChIP-Seq: 10ug per ChIP IP: 5ug per IP WB: 0.5-2ug/ml Note: Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt/Sonication Protocol when preparing nuclear extracts for Western blot. Researchers should empirically determine the suitability of the Lys7 polr2a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "RNA pol II CTD monomethyl Lys7, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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