17-680 ChIPAb+ Monomethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set

17-680
25 assays  25 assays per kit, ~2μg per chromatin immunoprecipitation
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      Vrt WB, ChIP-seq, ChIP
      Description
      Catalogue Number17-680
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Monomethyl-Histone H3 (Lys9) - 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+ Monomethyl-Histone H3 (Lys9) set includes the Anti-monomethyl-Histone H3 (Lys9) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers in the GAPDH coding region, amplifying a 213 base pair PCR product. The monomethyl-histone H3 (Lys9) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of monomethyl-histone H3 (Lys9) associated chromatin.
      Alternate Names
      • H3K9me1
      • Histone H3 (mono methyl K9)
      References
      Product Information
      FormatProtein G Purified
      Control
      • Included negative control mouse IgG antibody and control primers specific for human GAPDH coding region.
      PresentationAnti-monomethyl-Histone H3 (Lys9) (mouse monoclonal IgG2aқ, clone CMA306). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium azide. 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.

      ChIP Primers GAPDH Coding region. One vial containing 75 μL of 5 μM of each primer specific for a region of the human GAPDH coding region. Store at -20°C.
      FOR: GGC TCC CAC CTT TCT CAT CC
      REV: GGC CAT CCA CAG TCT TCT GG
      Quality LevelMQ100
      Applications
      ApplicationThis ChIPAb+ Validated Antibody & Primer Set conveniently includes the antibody, matched IgG negative control antibody & set control PCR primers that detect a known positive locus.
      Key Applications
      • Western Blotting
      • ChIP-seq
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from 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-monomethyl-Histone H3 (Lys9) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of monomethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using GAPDH Coding region primers versus Control Primers directed against the GAPDH promoter region (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 data of previous lot. HeLa acid extract (Lane 1) was resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-monomethyl Histone H3 (Lys9) (0.5 μg/mL).
      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 monomethyl-histone H3 (Lys9) purified antibody is made against a synthetic peptide (monomethylated at Lys9) corresponding to amino acids 1-18 of Histone H3.

      Epitopea.a. 1-18
      CloneCMA306
      HostMouse
      SpecificityRecognizes histone H3, Mr 17 kDa, monomethylated at Lys9.
      IsotypeIgG
      Species Reactivity
      • 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 WeightMonomethyl-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 HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-monomethyl-Histone H3 (Lys9) antibody and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of monomethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH Coding region (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+ Monomethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

      ChIPAb+ Monomethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 21471242147124
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 23910682391068
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 24245052424505
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 19579951957995
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 20367062036706
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 22119892211989
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 29528652952865
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 29725412972541
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 32903143290314
      ChIPAb+ Monomethyl-Histone H3 (Lys9) - 32924383292438

      References

      Reference overviewPub Med ID
      Prdm16 is required for the maintenance of brown adipocyte identity and function in adult mice.
      Harms, MJ; Ishibashi, J; Wang, W; Lim, HW; Goyama, S; Sato, T; Kurokawa, M; Won, KJ; Seale, P
      Cell metabolism  19  593-604  2014

      Show Abstract
      24703692 24703692
      The histone H3 methyltransferase G9A epigenetically activates the serine-glycine synthesis pathway to sustain cancer cell survival and proliferation.
      Ding, J; Li, T; Wang, X; Zhao, E; Choi, JH; Yang, L; Zha, Y; Dong, Z; Huang, S; Asara, JM; Cui, H; Ding, HF
      Cell metabolism  18  896-907  2013

      Show Abstract
      24315373 24315373

      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