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17-10241 ChIPAb+™ Acetyl-Histone H3 (Lys9/18) - ChIP Validated Antibody and Primer Set

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17-10241
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
      B, H, M, Yeast (S. cerevisiae)WB, ChIP
      Description
      Catalogue Number17-10241
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+™ Acetyl-Histone H3 (Lys9/18) - 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 (Lys9/18) set includes the Acetyl-Histone H3 (Lys9/18) antibody, a Normal rabbit serum, and control primers which amplify a 166 bp region of human GAPDH promoter. The Acetyl-Histone H3 (Lys9/18) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Acetyl-Histone H3 (Lys9/18)-associated chromatin.
      Alternate Names
      • H3K9/18Ac
      • Histone H3 (acetyl K9/18)
      Background InformationHistone H3 is one of the 5 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.

      Acetylation of histone H3 occurs at several different lysine positions in the histone tail and is performed by a family of enzymes known as Histone Acetyl Transferases (HATs).
      References
      Product Information
      FormatSerum
      Control
      • Includes normal rabbit serum and primers specific for human GAPDH promoter.
      PresentationAnti-Acetyl-Histone H3 (Lys9/18) (rabbit polyclonal), . One vial containing 25 µL of rabbit antiserum containing 0.05% sodium azide before the addition of glycerol to 30%. Store at -20°C.
      Normal Rabbit Serum, . One vial containing 25 μL of antiserum containing 0.05% sodium azide. Store at -20°C.
      Control Primers, human GAPDH promoter, . One vial containing 75 μL of 5 μM of each primer specific for human 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+ Acetyl-Histone H3 (Lys9/18) -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 Rabbit Serum, or 1 µL of Anti-Acetyl-Histone H3 (Lys9/18), and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of acetyl-Histone H3 (Lys9/18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus and human β-globin 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™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details
      Western Blot Analysis:
      Representative lot data.
      Acid extracts from untreated (lane 1), sodium butyrate treated (lane 2, Catalog # 17-305) HeLa cells and recombinant Histone
      H3, (Lane 3, Catalog # 14-411) were resolved by electrophoresis, transferred to nitrocellulose and probed with Anti-Acetyl-Histone H3 (Lys9/18) (1:5,000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
      Arrow indicates Acetyl-histone H3 (Lys9/18) (~17kDa). (Figure 3).
      Beadlyte® Histone-Peptide Specificity Assay:
      Representative lot data.
      1:1,000-1:40,000 dilutions of anti-acetyl-Histone H3 (Lys9/18) were incubated with a cocktail of microspheres conjugated to histone H3 peptides with the following modifications:
      1. acetyl-lysine 4
      2. acetyl-lysine 14
      3. acetyl-lysine 9
      4. acetyl-lysine 18
      5. No modifications, containing aa 1-20
      Unbound antibody was removed by filtration. The peptide-antibody complexes were incubated with a biotin-conjugated anti-mouse secondary antibody followed by incubation with a phycoerythrin-streptavidin conjugate.
      Fluorescence was read on a Luminex® 100™ instrument. Median
      fluorescence intensity (MFI) is plotted. (Figure 4).

      Biological Information
      ImmunogenKLH-conjugated synthetic peptide (..ARAcKSTGGKAPRAcKQL..) in which AcK corresponds to acetyl-lysine at residue 9 and 18 of human Histone H3.
      EpitopeAcetylated Lys9/18
      HostRabbit
      SpecificityRecognizes Histone H3 acetylated on lysines 9 and 18.
      Species Reactivity
      • Bovine
      • Human
      • Mouse
      • Yeast (S. cerevisiae)
      Species Reactivity NoteBroad species cross-reactivity expected
      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
      • MGC87783
      • H3.3A
      • MGC87782
      • H3F3
      • H3.3B
      • H3F3B
      Modifications
      • Acetylation
      Purification MethodUnpurified
      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:
      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 Rabbit Serum,or 1 µL of Anti-Acetyl-Histone H3 (Lys9/18) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Acetyl-Histone H3 (Lys9/18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus (Figure 1). Please refer to the EZ-Magna ChIP™ A (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. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
      Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
      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+™ Acetyl-Histone H3 (Lys9/18) - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ Acetyl-Histone H3 (Lys9/18) - 1992289 1992289
      ChIPAb+ Acetyl-Histone H3 (Lys9/18) - 2053034 2053034
      ChIPAb+ Acetyl-Histone H3 (Lys9/18) - NRG1880394 NRG1880394
      ChIPAb+™ Acetyl-Histone H3 (Lys9/18) 3171005
      ChIPAb+™ Acetyl-Histone H3 (Lys9/18) Polyclonal Antibody/Primer Set 3766912
      ChIPAb+™ Acetyl-Histone H3 (Lys9/18) Polyclonal Antibody/Primer Set 2965346
      ChIPAb+™ Acetyl-Histone H3 (Lys9/18) Polyclonal Antibody/Primer Set 2869079
      ChIPAb+™ Acetyl-Histone H3, (Lys9/18), Polyclonal Antibody/Primer Set 2837839

      References

      Reference overviewPub Med ID
      An ARID domain-containing protein within nuclear bodies is required for sperm cell formation in Arabidopsis thaliana.
      Zheng, B; He, H; Zheng, Y; Wu, W; McCormick, S
      PLoS genetics  10  e1004421  2014

      Show Abstract
      25057814 25057814

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      Title
      Advance your Epigenetics Research

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      Categories

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