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05-745R-S Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Trial Size

Use Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Recombinant, Trial Size (rabbit monoclonal antibody) proven in WB, ChIP, ChIP-seq, ELISA, PIA, Mplex, DB to detect trimethyl-Histone H3

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Research Category: Epigenetics & Nuclear Function Research Sub-Category: Histones Gene Symbol: H3F3A, H3F3B UniProt Number: Q16695
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05-745R-S
10 µL  
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Key Spec Table

Species ReactivityKey ApplicationsHostFormatAntibody Type
HWB, ChIP, ChIP-seq, DB, ELISA, PIA, MplexRbPurifiedMonoclonal Antibody
Description
Catalogue Number05-745R-S
Replaces05-745
Trade Name
  • Upstate
DescriptionAnti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Trial Size
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.
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.
Quality LevelMQ100
Applications
ApplicationUse Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Recombinant, Trial Size (rabbit monoclonal antibody) proven in WB, ChIP, ChIP-seq, ELISA, PIA, Mplex, DB to detect trimethyl-Histone H3 (Lys4) also known as H3K4me3.
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-S), 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
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 Size10 µL
Global Trade Item Number
Catalogue Number GTIN
05-745R-S 04053252946844