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17-10242 ChIPAb+™ Trimethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set

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17-10242
25 assays  25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, VrtWB, ChIP
      Description
      Catalogue Number17-10242
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+™ Trimethyl-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+ Trimethyl-Histone H3 (Lys9) set includes the Trimethyl-Histone H3 (Lys9) antibody, a Normal mouse IgG, and control primers which amplify a 117 bp region of ChIP Primers, ZNF554. The Trimethyl-Histone H3 (Lys9) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Trimethyl-Histone H3 (Lys9) -associated chromatin.
      Alternate Names
      • H3K9me3
      • Histone H3 (tri methyl K9)
      • H3 histone family, member T
      • histone 3, H3
      • histone cluster 3, H3
      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. Histone proteins are highly post-translationally modified however Histone H3 is the most extensively modified of the five histones. Histone H3 sequence variants and variable modification states are thought to play a role in the dynamic and long term regulation of genes. Trimethylation of histone H3 on Lys9 (H3K9me3) is one of the most highly studied epigenetic marks. H3K9me3 functions in the repression of euchromatic genes, and in epigenetic control of heterochromatin assembly, most likely by acting as a recognition motif for the binding of chromatin-associated proteins, such as Swi6 or HP1α. The enzymes responsible for H3K9 trimethylation are SUV39H1 and SUV39H2.
      References
      Product Information
      FormatPurified
      Control
      • Includes normal mouse IgG and primers specific for human ZNF554.
      PresentationAnti-Trimethyl-Histone H3 (Lys9) (mouse monoclonal). One vial containing 25 µL of purified mouse monoclonal IgG1 in PBS with 0.05% sodium azide before the addition of glycerol to 30%. Store at -20°C.
      Normal Mouse IgG. One vial containing 25 µg of purified IgG in 25 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.
      Control Primers, ZNF554. One vial containing 75 μL of 5 μM of each primer specific for the 3’ end of human ZNF554. Store at -20°C.
      FOR: CGG GGA AAA GCC CTA TAA AT
      REV: TCC ACA TTC ACT GCA TTC GT
      Quality LevelMQ100
      Applications
      ApplicationThis ChIPAb+ Trimethyl-Histone H3 (Lys9) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Representative lot data.
      Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Mouse IgG,or 1 µL of Anti-Trimethyl Histone H3 (Lys9), and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of trimethyl Histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP primers specific for the human ZNF554 region as a positive locus and human GAPDH primers as a negative locus. (Figure 2). 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 ChIP™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
      Western Blot Analysis:
      Representative lot data.
      Recombinant Histone H3 (Cat. #14-494) (Lane 1) and HeLa acid extract (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-trimethyl Histone H3 (Lys9), clone CMA308 (2 μg/mL). Proteins were visualized using
      a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
      Arrow indicates trimethyl Histone H3 (Lys9) (~17 kDa). (Figure 3).
      Biological Information
      ImmunogenKLH-conjugated synthetic peptide corresponding to human Histone H3 trimethylated on Lys9.
      EpitopeTrimethyl Lys9
      CloneCMA308
      HostMouse
      SpecificityThis antibody recognizes Histone H3 trimethylated at Lys9. Some cross-reactivity with dimethyl Lys9 is detected at higher concentrations, but the difference in specificity is >27-fold1. Phosphorylation of Ser10 interferes with antibody binding to trimethyl Lys91.
      IsotypeIgG1κ
      Species Reactivity
      • Human
      • Vertebrates
      Species Reactivity NoteHuman. Broad species cross-reactivity is expected, based on 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. 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. [provided by RefSeq]
      Gene Symbol
      • H3.
      • H3/g
      • H3/t
      • H3FT
      • H3T
      • H3t
      • MGC126886
      • MGC126888
      • OTTHUMP00000037945
      Modifications
      • Methylation
      Purification MethodProtein G Purified
      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 Weight~17 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Representative lot data.
      Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Mouse IgG, or 1 µL of Anti-Trimethyl Histone H3(Lys9), and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of trimethyl Histone H3 (Lys9) associated DNA fragments was verified by qPCR using ChIP primers specific for the human ZNF554 region as a positive locus(Figure 1). Please refer to the EZ-Magna ChIP™ G (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 2-8°C from date of receipt. For maximum recovery of product, centrifuge the vial prior to removing the cap. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+™ Trimethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

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

      TitleLot Number
      ChIPAb+ Trimethyl-Histone H3 (Lys9) - 1990445 1990445
      ChIPAb+ Trimethyl-Histone H3 (Lys9) - 2074786 2074786
      ChIPAb+ Trimethyl-Histone H3 (Lys9) - 2167190 2167190
      ChIPAb+ Trimethyl-Histone H3 (Lys9) - NRG1889589 NRG1889589
      ChIPAb+™ Trimethyl-Histone H3 (Lys9) Monoclonal Antibody/Primer Set 2982578
      ChIPAb+™ Trimethyl-Histone H3 (Lys9) Monoclonal Antibody/Primer Set 2978319
      ChIPAb+™ Trimethyl-Histone H3 (Lys9) Monoclonal Antibody/Primer Set 2874538
      ChIPAb+™ Trimethyl-Histone H3 -2552614 2552614
      ChIPAb+™ Trimethyl-Histone H3 -2794083 2794083
      ChIPAb+™ Trimethyl-Histone H3 (Lys9) - 2449242 2449242

      Brochure

      Title
      Advance your Epigenetics Research (MilliporeSigma)

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      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