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17-678 ChIPAb+ Trimethyl-Histone H3 (Lys4) - ChIP Validated Antibody and Primer Set

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17-678
25 assays  25 assays per kit, ~2μg per chromatin immunoprecipitation
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, VrtWB, ICC, ChIP, ChIP-seq
      Description
      Catalogue Number17-678
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Trimethyl-Histone H3 (Lys4) - ChIP Validated Antibody and Primer Set
      OverviewAll 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+ Trimethyl-Histone H3 (Lys4) set includes the Anti-trimethyl-Histone H3 (Lys4) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 166 bp region within the promoter of the human GAPDH gene. The trimethyl-Histone H3 (Lys4) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of trimethyl-histone H3 (Lys4)-associated chromatin.
      Alternate Names
      • H3K4me3
      • Histone H3 (tri methyl K4)
      References
      Product Information
      FormatProtein G Purified
      Control
      • Included negative control mouse IgG antibody and control primers specific for human GAPDH promoter.
      PresentationAnti-trimethyl-Histone H3 (Lys4) (mouse monoclonal IgG1, Clone CMA304). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium. Store at -20°C.

      Normal Mouse IgG. Two vials containing 25 μg purified mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.

      Control Primers. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of GAPDH. Store at -20°C.
      FOR: TAC TAG CGG TTT TAC GGG CG
      REV: TCG AAC AGG AGG AGC AGA GAG CGA

      Quality LevelMQ100
      Applications
      ApplicationThis ChIPAb+ Trimethyl-Histone H3 (Lys4) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Immunocytochemistry
      • Chromatin Immunoprecipitation (ChIP)
      • ChIP-seq
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from untreated or colcemid treated HeLa cells (1 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-trimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of trimethyl-Histone H3 (Lys4) associated DNA fragments was verified by qPCR using primers ampliflying a region of the human B-Globin promoter or using GAPDH promoter 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. #17-408) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.

      Western Blot Analysis:
      Representative lot data.
      HeLa acid extract were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-trimethyl Histone H3 (Lys4), clone CMA304 at 1 μg/ml (lane 1) or 0.5 μg/ml (lane 2). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP (Cat. #AP124P) and a chemiluminescence detection system (Please see figures).
      Biological Information
      ImmunogenThe trimethyl-histone H3 (Lys4) purified antibody is made against a synthetic peptide (trimethylated at Lys4) corresponding to amino acids 1-12 of human histone H3.
      Epitopea.a. 1-12
      CloneCMA304
      HostMouse
      SpecificityRecognizes histone H3, Mr 17 kDa, trimethylated at Lys4.
      IsotypeIgG1κ
      Species Reactivity
      • Human
      • Vertebrates
      Species Reactivity NoteHuman. The immunogen sequence is identical in a wide range of animal and plant species.
      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. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3.
      Modifications
      • Methylation
      UniProt Number
      UniProt SummaryFUNCTION:Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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. Ref.14 Ref.18 Ref.22

      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. Interacts with HIRA, a chaperone required for its incorporation into nucleosomes. Ref.14

      SUBCELLULAR LOCATION: Nucleus.

      Developmental stage Expressed throughout the cell cycle independently of DNA synthesis.

      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.

      Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription.

      Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression.

      Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5, Lys-37 and Lys-80. 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, which are linked to gene repression, are underrepresented. 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.

      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, probably DAPK3. Phosphorylation at 'Ser-11' by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation on Ser-32 is specific to regions bordering centromeres in metaphase chromosomes. Ref.9 Ref.10 Ref.12 Ref.13 Ref.19 Ref.20 Ref.21 Ref.29

      Ubiquitinated By similarity.

      SIMILARITY: Belongs to the histone H3 family.

      SEQUENCE CAUTION: The sequence CAH73371.1 differs from that shown. Reason: Erroneous gene model prediction.

      Molecular WeightTrimethyl histone H3 at ~17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from colcemid-reated HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either normal mouse IgG or Anti-trimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immuno-precipitation of trimethyl-histone H3 (Lys4) associated DNA fragments was verified by qPCR using Control Primers (Please see figures).
      Please refer to the EZ-Magna G ChIP™ (Cat. #17-409) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.
      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. Aliquot upon thawing, avoid freeze thaw cycles.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per kit, ~2μg per chromatin immunoprecipitation
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+ Trimethyl-Histone H3 (Lys4) - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

      ChIPAb+ Trimethyl-Histone H3 (Lys4) - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ Trimethyl-Histone H3 (Lys4) - 2137056 2137056
      ChIPAb+ Trimethyl-Histone H3 (Lys4) - JH1752509 JH1752509
      ChIPAb+ Trimethyl-Histone H3 (Lys4) - NG1834921 NG1834921
      ChIPAb+ Trimethyl-Histone H3 (Lys4) - NG1932156 NG1932156
      ChIPAb+ Trimethyl-Histone H3 (Lys4) - NG1947170 NG1947170
      ChIPAb+ Trimethyl-Histone H3 (Lys4) - NRG1583029 NRG1583029
      ChIPAb+™ Trimethyl-Histone H3 (Lys4) 2470946
      ChIPAb+™ Trimethyl-Histone H3 (Lys4) - 3452500 3452500
      ChIPAb+™ Trimethyl-Histone H3 (Lys4) - 3777360 3777360
      ChIPAb+™ Trimethyl-Histone H3 (Lys4) -2705061 2705061

      References

      Reference overviewPub Med ID
      Alterations of epigenetic signatures in hepatocyte nuclear factor 4α deficient mouse liver determined by improved ChIP-qPCR and (h)MeDIP-qPCR assays.
      Zhang, Q; Lei, X; Lu, H
      PloS one  9  e84925  2014

      Show Abstract
      24427299 24427299
      Deletion of a conserved cis-element in the Ifng locus highlights the role of acute histone acetylation in modulating inducible gene transcription.
      Balasubramani, A; Winstead, CJ; Turner, H; Janowski, KM; Harbour, SN; Shibata, Y; Crawford, GE; Hatton, RD; Weaver, CT
      PLoS genetics  10  e1003969  2014

      Show Abstract
      24415943 24415943
      The gene signature in CCAAT-enhancer-binding protein α dysfunctional acute myeloid leukemia predicts responsiveness to histone deacetylase inhibitors.
      Liss, A; Ooi, CH; Zjablovskaja, P; Benoukraf, T; Radomska, HS; Ju, C; Wu, M; Balastik, M; Delwel, R; Brdicka, T; Tan, P; Tenen, DG; Alberich-Jorda, M
      Haematologica  99  697-705  2014

      Show Abstract
      24162792 24162792
      Genome-wide evaluation of histone methylation changes associated with leaf senescence in Arabidopsis.
      Brusslan, JA; Rus Alvarez-Canterbury, AM; Nair, NU; Rice, JC; Hitchler, MJ; Pellegrini, M
      PloS one  7  e33151  2012

      Show Abstract
      22427974 22427974

      Brochure

      Title
      Advance your Epigenetics Research (MilliporeSigma)

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      Product Families

      Categories

      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Primary Antibodies
      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies