17-648 ChIPAb+ Dimethyl-Histone H3 (Lys9) - ChIP Validated Antibody and Primer Set

17-648
25 assays  25 assays per kit, ~4μL per chromatin immunoprecipitation
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, M, Ma WB, IP, ChIP
      Description
      Catalogue Number17-648
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Dimethyl-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+ Dimethyl-Histone H3 (Lys9) set includes the anti-dimethyl-histone H3 (Lys9) antibody, a negative control antibody (normal rabbit serum), and qPCR primers which amplify a 110 bp region within the promoter of the human β-globin gene. The dimethyl-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 dimethyl-histone H3 (Lys9) associated chromatin.
      Alternate Names
      • H3K9me2
      • Histone H3 (di methyl K9)
      • Histone H3K9me2
      Background InformationThe methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 9 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SuvH39H1 or G9a. This methylated lysine can be demethylated by histone demethylases as JMJD1A, LSD1 or JMJD2C. Methylation of this residue is mainly associated with transcriptional repression.
      References
      Product Information
      Components
      • Anti-Dimethyl-Histone H3 (Lys9) (rabbit serum), 1 vial
      • Negative ChIP Control serum, 1 vial
      • ChIP Primers β-globin , 1 vial
      FormatSerum
      Control
      • Included negative control antibody normal rabbit serum and control primers specific for human β-globin promoter.
      PresentationDimethyl-Histone H3 (Lys9) (rabbit polyclonal serum). One vial containing 100 μL of antiserum containing 0.05% sodium azide.

      Normal Rabbit Serum. One vial containing 100 uL antiserum containing 0.05% sodium azide.

      ChIP Primers, β-Globin. One vial containing 75 μL of 5 μM of each primer specific for human β-globin.
      FOR: AGG ACA GGT ACG GCT GTC ATC
      REV: TTT ATG CCC AGC CCT GGC TC
      Quality LevelMQ100
      Applications
      ApplicationDimethyl-Histone H3 (Lys9) ChIP validated antibody & primer set including the ChIP-grade antibody & the specific control PCR primers used for chromatin immunoprecipitation of H3K9Me2.
      Key Applications
      • Western Blotting
      • Immunoprecipitation
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents) was subjected to chromatin Immunoprecipitation using 4 μL of either a normal rabbit antiserum or Antidimethyl-Histone H3 (Lys9) serum and the Magna ChIP A (Cat. #17-610) Kit (Please see figures). Successful Immunoprecipitation of dimethyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using β-globin ChIP Primers flanking the human β-globin promoter or primers amplifying the promoter of human GAPDH, which is transcriptionally inactive in HeLa cells. Percent Input relative to standard curves for each qPCR primer set are shown.
      Please refer to the EZ-Magna A ChIP™ (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

      Western blot analysis and peptide inhibition:
      HeLa Acid extract were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-dimethyl-Histone H3 (Lys9) (1:500, Lane 1) or preincubated with 0.4 μM Histone H3 peptide with following modifications:
      Lane 2: Linear non-modified
      Lane 3: Branched non-modified
      Lane 4: Branched trimethyl
      Lane 5: Linear trimethyl
      Lane 6: Branched dimethyl
      Lane 7: Linear dimethyl
      Lane 8: Branched monomethyl
      Lane 9: Linear monomethyl
      Proteins were visualized using a goat anti–rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
      Biological Information
      ImmunogenThe dimethyl-histone H3 (Lys9) rabbit serum is made against a KLH-conjugated, branched synthetic peptide containing the sequence ..Rme2KSTG.., in which me2K corresponds to dimethyl-lysine at residue 9 of human histone H3
      EpitopeDimethyl Lys9
      HostRabbit
      SpecificityDimethyl-Histone H3 (Lys9)
      Species Reactivity
      • Human
      • Mouse
      • Mammals
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene contains introns and its mRNA is polyadenylated, unlike most histone genes. The protein encoded is a replication-independent member of the histone H3 family.
      Gene Symbol
      • H3F3A
      • H3F3B
      • H3F3
      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.
      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 AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents) was subjected to chromatin immunoprecipitation using 4 μL of either a normal rabbit antiserum or 4 μL Anti-Dimethyl-Histone H3 (Lys9) serum and the Magna ChIP A (Cat. #17-610) Kit.
      Successful immunoprecipitation of dimethyl histone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the β-globin human promoter (Please see figures).
      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
      Packaging Information
      Material Size25 assays
      Material Package25 assays per kit, ~4μL per chromatin immunoprecipitation
      Transport Information
      Supplemental Information
      Specifications

      Documentation

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

      Title

      Safety Data Sheet (SDS) 

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

      TitleLot Number
      ChIPAb+ Dimethyl-Histone H3 (Lys9) -28033112803311
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum2483961
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum - 21366402136640
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum - 23835992383599
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum - 24556592455659
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum - 19975011997501
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum - 20347262034726
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum - 21933312193331
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum - 22935292293529
      ChIPAb+ Dimethyl-Histone H3 (Lys9) Serum - 28149612814961

      References

      Reference overviewApplicationSpeciesPub Med ID
      ZEB1-associated drug resistance in cancer cells is reversed by the class I HDAC inhibitor mocetinostat.
      Meidhof, S; Brabletz, S; Lehmann, W; Preca, BT; Mock, K; Ruh, M; Schüler, J; Berthold, M; Weber, A; Burk, U; Lübbert, M; Puhr, M; Culig, Z; Wellner, U; Keck, T; Bronsert, P; Küsters, S; Hopt, UT; Stemmler, MP; Brabletz, T
      EMBO molecular medicine  7  831-47  2015

      Show Abstract
      25872941 25872941
      Nuclear actin filaments recruit cofilin and actin-related protein 3, and their formation is connected with a mitotic block.
      Kalendová, A; Kalasová, I; Yamazaki, S; Uličná, L; Harata, M; Hozák, P
      Histochemistry and cell biology  142  139-52  2014

      Show Abstract
      25002125 25002125
      Matrix softness regulates plasticity of tumour-repopulating cells via H3K9 demethylation and Sox2 expression.
      Tan, Y; Tajik, A; Chen, J; Jia, Q; Chowdhury, F; Wang, L; Chen, J; Zhang, S; Hong, Y; Yi, H; Wu, DC; Zhang, Y; Wei, F; Poh, YC; Seong, J; Singh, R; Lin, LJ; Doğanay, S; Li, Y; Jia, H; Ha, T; Wang, Y; Huang, B; Wang, N
      Nature communications  5  4619  2014

      Show Abstract
      Western BlottingMouse25099074 25099074
      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
      Histone H3K9 methyltransferase G9a represses PPARγ expression and adipogenesis.
      Wang, L; Xu, S; Lee, JE; Baldridge, A; Grullon, S; Peng, W; Ge, K
      The EMBO journal  32  45-59  2013

      Show Abstract
      Western Blotting23178591 23178591
      Expression and epigenetic regulation of angiogenesis-related factors during dormancy and recurrent growth of ovarian carcinoma.
      Lyu, T; Jia, N; Wang, J; Yan, X; Yu, Y; Lu, Z; Bast, RC; Hua, K; Feng, W
      Epigenetics  8  1330-46  2013

      Show Abstract
      24135786 24135786
      Antisilencing role of the RNA-directed DNA methylation pathway and a histone acetyltransferase in Arabidopsis.
      Li, X; Qian, W; Zhao, Y; Wang, C; Shen, J; Zhu, JK; Gong, Z
      Proceedings of the National Academy of Sciences of the United States of America  109  11425-30  2012

      Show Abstract
      Western Blotting22733760 22733760
      5-Aza-2'-deoxycytidine reactivates gene expression via degradation of pRb pocket proteins.
      Zheng, Z; Li, L; Liu, X; Wang, D; Tu, B; Wang, L; Wang, H; Zhu, WG
      FASEB journal : official publication of the Federation of American Societies for Experimental Biology  26  449-59  2012

      Show Abstract
      21990374 21990374
      Zinc-induced Dnmt1 expression involves antagonism between MTF-1 and nuclear receptor SHP.
      Zhang, Y; Andrews, GK; Wang, L
      Nucleic acids research  40  4850-60  2012

      Show Abstract
      22362755 22362755
      Plant 45S rDNA clusters are fragile sites and their instability is associated with epigenetic alterations.
      Huang, M; Li, H; Zhang, L; Gao, F; Wang, P; Hu, Y; Yan, S; Zhao, L; Zhang, Q; Tan, J; Liu, X; He, S; Li, L
      PloS one  7  e35139  2012

      Show Abstract
      22509394 22509394

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      Advance your Epigenetics Research (MilliporeSigma)

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