05-745R Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, rabbit monoclonal

05-745R
100 µL  
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H WB, ChIP, ChIP-seq, DB, ELISA, PIA, Mplex Rb Purified Monoclonal Antibody
      Description
      Catalogue Number05-745R
      Replaces05-745
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionAnti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, rabbit monoclonal
      OverviewRecombinant version of Cat. # 05-745, clone MC315
      Alternate Names
      • H3K4me3
      • Histone H3 (tri methyl K4)
      Background InformationHistone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure.

      The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.
      References
      Product Information
      FormatPurified
      PresentationProtein A purified, recombinant antibody in 60 mM Tris, 90 mM NaCl, 0.03% NaN3, 60 mM Glycine, pH 7.5, 40% Glycerol.
      Applications
      ApplicationUse Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Recombinant (rabbit monoclonal antibody) proven in WB, ChIP, ChIP-seq, ELISA, PIA, Mplex, DB to detect trimethyl-Histone H3 (Lys4) also known as H3K4me3, Histone H3 (tri methyl K4).
      Key Applications
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      • ChIP-seq
      • Dot Blot
      • ELISA
      • Peptide Inhibition Assay
      • Multiplexing
      Application NotesChIP-seq Analysis:
      Representative lot data. Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 3 µL of Anti-trimethyl-Histone H3 (Lys4) antibody (cat# 05-745R), 20 µL Protein A/G beads, and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of eighteen million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files. The highest 25% of peaks identified in the 05-745R and 07-473 datasets showed 99% overlap with peaks identified in the ENCODE H3K4me3 BROAD Histone track for HeLa S3.
      Biological Information
      ImmunogenBSA-conjugated, synthetic peptide containing the sequence …RTme3KQT… in which me3K corresponds to trimethyl-lysine 4 of human histone H3.
      Clone15-10C-E4
      HostRabbit
      SpecificityRecognizes Histone H3 containing trimethyl-lysine 4 and, to a lesser extent, dimethyl-lysine 4. See certificate of analysis for further details.
      IsotypeIgG
      Species Reactivity
      • Human
      Species Reactivity NoteBroad species cross-reactivity expected due to sequence homology
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures.
      Gene Symbol
      • H3F3A
      • H3F3B
      Modifications
      • Methylation
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q16695 # Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
      SIZE: 136 amino acids; 15508 Da
      SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA.
      SUBCELLULAR LOCATION: Nucleus.
      TISSUE SPECIFICITY: Expressed in testicular cells.
      DEVELOPMENTAL STAGE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
      PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity).
      SIMILARITY: SwissProt: Q16695 ## Belongs to the histone H3 family.
      Molecular Weight17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely evaluated by immunoblot on HeLa nuclear extract
      Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at -20°C from date of receipt.
      Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, rabbit monoclonal SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, rabbit monoclonal Certificates of Analysis

      TitleLot Number
      Anti-trimethyl-Histone H3 (Lys4), clone -25591722559172
      Anti-trimethyl-Histone H3 (Lys4), clone -27133272713327
      Anti-trimethyl-Histone H3 (Lys4), clone -28138672813867
      Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4 -31580713158071
      Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant - 23904302390430
      Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant - 24204052420405
      Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant - 24333762433376
      Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant - 19636121963612
      Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant - 20231972023197
      Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant - 21530872153087

      References

      Reference overviewApplicationPub Med ID
      A dual role for SAGA-associated factor 29 (SGF29) in ER stress survival by coordination of both histone H3 acetylation and histone H3 lysine-4 trimethylation.
      Schram, AW; Baas, R; Jansen, PW; Riss, A; Tora, L; Vermeulen, M; Timmers, HT
      PloS one  8  e70035  2013

      Show Abstract
      23894581 23894581
      Chromatin signatures at transcriptional start sites separate two equally populated yet distinct classes of intergenic long noncoding RNAs.
      Marques, AC; Hughes, J; Graham, B; Kowalczyk, MS; Higgs, DR; Ponting, CP
      Genome biology  14  R131  2013

      Show Abstract
      24289259 24289259
      An efficient immunodetection method for histone modifications in plants.
      Nic-Can, G; Hernández-Castellano, S; Kú-González, A; Loyola-Vargas, VM; De-la-Peña, C
      Plant methods  9  47  2013

      Show Abstract
      24341414 24341414
      Coordinate H3K9 and DNA methylation silencing of ZNFs in toxicant-induced malignant transformation.
      Severson, PL; Tokar, EJ; Vrba, L; Waalkes, MP; Futscher, BW
      Epigenetics  8  1080-8  2013

      Show Abstract
      23974009 23974009
      The Drosophila melanogaster CHD1 chromatin remodeling factor modulates global chromosome structure and counteracts HP1a and H3K9me2.
      Bugga, L; McDaniel, IE; Engie, L; Armstrong, JA
      PloS one  8  e59496  2013

      Show Abstract
      Immunofluorescence23533627 23533627
      A map of the cis-regulatory sequences in the mouse genome.
      Shen, Yin, et al.
      Nature, 488: 116-20 (2012)  2012

      Show Abstract
      22763441 22763441
      Dynamics and memory of heterochromatin in living cells.
      Hathaway, NA; Bell, O; Hodges, C; Miller, EL; Neel, DS; Crabtree, GR
      Cell  149  1447-60  2012

      Show Abstract
      22704655 22704655
      Histone crosstalk directed by H2B ubiquitination is required for chromatin boundary integrity.
      Ma, MK; Heath, C; Hair, A; West, AG
      PLoS genetics  7  e1002175  2011

      Show Abstract
      21811414 21811414
      Arabidopsis COMPASS-like complexes mediate histone H3 lysine-4 trimethylation to control floral transition and plant development.
      Jiang, D; Kong, NC; Gu, X; Li, Z; He, Y
      PLoS genetics  7  e1001330  2011

      Show Abstract
      Western Blotting21423667 21423667
      Polycomb eviction as a new distant enhancer function.
      Vernimmen, D; Lynch, MD; De Gobbi, M; Garrick, D; Sharpe, JA; Sloane-Stanley, JA; Smith, AJ; Higgs, DR
      Genes & development  25  1583-8  2011

      Show Abstract
      21828268 21828268

      Technical Info

      Title
      Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4 ChIP-seq Analysis
      White Paper - The Message in the Marks: Deciphering Cancer Epigenetics (EMD)

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      Accessories

      Catalog Number Description  
      04-745 Anti-trimethyl-Histone H3 (Lys4) Antibody, clone MC315, rabbit monoclonal Show Pricing & Availability

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      Life Science Research > Antibodies and Assays > Primary Antibodies