17-609 ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - ChIP Validated Antibody and Primer Set

17-609
25 assays  25 assays per kit, ~1μL per chromatin immunoprecipitation
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, M, Ma WB, ChIP
      Description
      Catalogue Number17-609
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ Acetyl-Histone H3 (Lys9) Serum - 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+ Acetyl-Histone H3 (Lys9) set includes the anti-acetyl-histone H3 (Lys9) antibody, a negative control antibody (normal rabbit serum), and qPCR primers which amplify a 166 base pair region within the promoter of the human GAPDH gene. The acetyl-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 acetyl-histone H3 (Lys9) associated chromatin.
      Alternate Names
      • H3K9Ac
      • Histone H3 (acetyl K9)
      References
      Product Information
      FormatSerum
      Control
      • Included negative control antibody normal rabbit serum and control primers specific for human GAPDH promoter.
      PresentationAnti-Acetyl-Histone H3 (Lys9) (rabbit polyclonal serum). One vial containing 25 μL of antiserum containing 0.05% sodium azide before the addition of glycerol to 30%.

      Normal Rabbit Serum. One vial containing 25 uL antiserum containing 0.05% sodium azide.

      Control Primers. One vial containing 75 μL of 5 μM of each primer specific for for human GAPDH.
      FOR: TAC TAG CGG TTT TAC GGG CG
      REV: TCG AAC AGG AGG AGC AGA GAG CGA

      Applications
      ApplicationThis ChIPAb+ Acetyl-Histone H3 (Lys9) Serum -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from untreated HeLa cells (1 X 106 cell equivalents) was subjected to chromatin immunoprecipitation using 1 μL of either a normal rabbit antiserum or Anti-Acetyl-Histone H3 (Lys9) serum and the Magna ChIP A Kit (Cat. #17-610) (Figure 2). Successful immunoprecipitation of acetyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter or primers amplifying the promoter of human β-globin, which is transcriptionally inactive in HeLa cells. Percent Input relative to standard curves for each qPCR primer set are shown, with immunoprecipitated DNA from control serum shown as (-) and acetylhistone H3 serum as (+).
      Please refer to the EZ-Magna A ChIP™ (Cat. #17-408) or EZ-ChIP™ (Cat. #17-371) protocol for experimental details.

      Western Blot Analysis:
      Acid-extracted proteins from normal HeLa cells (Lane 1) and HeLa cells treated with 5mM sodium butyrate for 24 hours (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-Acetyl-Histone H3 (Lys9) (1:5000). Proteins were visualized using a goat-anti rabbit secondary antibody conjugated to HRP and chemiluminescent detection system (Please see figures).
      Biological Information
      ImmunogenThe acetyl-histone H3 (Lys9) antiserum is made against a peptide corresponding to amino acids 4-14 of yeast histone H3 acetylated on lysine 9.
      HostRabbit
      SpecificityAcetyl-Histone H3 (Lys9)
      Species Reactivity
      • Human
      • Mouse
      • Mammals
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • H3F3A
      • H3F3B
      • H3F3
      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 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 1 μL of either a normal rabbit antiserum or Anti-Acetyl-Histone H3 (Lys9) serum and the Magna ChIP A (Cat. #17-610) Kit.
      Successful immunoprecipitation of acetylhistone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter (Please see figures).
      Please refer to the EZ-Magna A ChIP™ (Cat. #17-408) 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
      Packaging Information
      Material Size25 assays
      Material Package25 assays per kit, ~1μL per chromatin immunoprecipitation
      Transport Information
      Supplemental Information
      Specifications

      Documentation

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

      Title

      Safety Data Sheet (SDS) 

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

      TitleLot Number
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - 24595642459564
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - 21596682159668
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - 22088062208806
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - 31667493166749
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - 32589473258947
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - 33269093326909
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - 34552543455254
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - RF0807003RF0807003
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - RF0807003ARF0807003A
      ChIPAb+ Acetyl-Histone H3 (Lys9) Serum - RF0807003BRF0807003B

      References

      Reference overviewApplicationPub Med ID
      Epigenetic suppression of mouse Per2 expression in the suprachiasmatic nucleus by the inhalational anesthetic, sevoflurane.
      Mori, K; Iijima, N; Higo, S; Aikawa, S; Matsuo, I; Takumi, K; Sakamoto, A; Ozawa, H
      PLoS One  9  e87319  2014

      Show Abstract
      24498074 24498074
      Polyamine analogs modulate gene expression by inhibiting lysine-specific demethylase 1 (LSD1) and altering chromatin structure in human breast cancer cells.
      Zhu, Q; Huang, Y; Marton, LJ; Woster, PM; Davidson, NE; Casero, RA
      Amino Acids  42  887-98  2012

      Show Abstract
      Western Blotting21805138 21805138

      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