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17-10111 ChIPAb+™ Acetyl-Histone H3 (Lys18) - ChIP Validated Antibody and Primer Set

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17-10111
25 assays  25 assays per set. Recommended use: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, Vrt, Yeast (S. cerevisiae)WB, DB, ChIP, ChIP-seq
      Description
      Catalogue Number17-10111
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+™ Acetyl-Histone H3 (Lys18) - 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 (Lys18) set includes the Acetyl-Histone H3 (Lys18) antibody, a negative control normal rabbit serum, and control primers which amplify a 166 bp region of human GAPDH promoter. The Acetyl-Histone H3 (Lys18) 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 (Lys18) -associated chromatin.
      Alternate Names
      • H3K18Ac
      • Histone H3 (acetyl K18)
      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 negative control normal rabbit serum and primers specific for human GAPDH promoter.
      PresentationAnti-Acetyl-Histone H3 (Lys18) (rabbit polyclonal). One vial of 50 µL of rabbit serum containing 0.05% sodium azide and 30% glycerol. Store at -20°C.
      Normal Rabbit Serum. One vial of 100 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 each of primer specific for human GAPDH promoter. 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 (Lys18) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Western Blotting
      • Dot Blot
      • Chromatin Immunoprecipitation (ChIP)
      • ChIP-seq
      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 2 µL of either Normal Rabbit Serum , or 2 µL of Anti-Acetyl-Histone H3 (Lys18)and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Acetyl-Histone H3 (Lys18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region as a positive locus, and β-globin primers as a negative locus. 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.

      ChIP-Sequencing:
      Representative lot data.
      Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 2 µg of anti-acetyl-Histone H3 (Lys18) antibody (cat# 17-10111), 20 µL Protein A/G beads , and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq™ instrument. An excess of ten million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files.

      Western Blot Analysis:
      Representative lot data.
      Recombinant histone H3 (lane 1, Catalog # 14-494) and acid extracts from sodium butyrate treated (lane 2) and untreated (lane 3) HeLa cells (Catalog # 17-305) were probed with anti acetyl- Histone H3 (Lys18) (1:10,000 dilution).
      Arrow indicates acetyl histone H3 (Lys18) (17 kDa).

      Dot Blot:
      Representative lot data.
      40 ng and 4ng amounts of histone peptides with various modifications (see table 1) were transferred to PVDF membrane and probed with Anti-Acetyl-Histone H3 (Lys18) antibody (1:5000 dilution). Proteins were visualized using a goat anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system. Image from a 60 second exposure is shown.
      Biological Information
      ImmunogenKLH-conjugated, synthetic peptide (GKAPRAcKQLASK-C) corresponding to amino acids 13-23 of yeast Histone H3 acetylated on lysine 18with a C-terminal cysteine added for conjugation purposes
      HostRabbit
      SpecificityRecognizes Histone H3 acetylated on lysine 18.
      Species Reactivity
      • Human
      • Vertebrates
      • Yeast (S. cerevisiae)
      Species Reactivity NoteYeast, Human, Most common vertebrates
      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. 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.
      Gene Symbol
      • HIST3H3
      • H3FT
      • MGC126886
      • H3t
      • MGC126888
      • H3T
      • H3/g
      • H3.4
      • H3/t
      Modifications
      • Acetylation
      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 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 2 µL of either Normal Rabbit Serum, or 2 µL of Anti-Acetyl-Histone H3 (Lys18)and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Acetyl-Histone H3 (Lys18) associated DNA fragments was verified by qPCR using Control Primers specific for the human GAPDH promoter region (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: ~2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+™ Acetyl-Histone H3 (Lys18) - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ Acetyl-Histone H3 (Lys18) - 2049804 2049804
      ChIPAb+ Acetyl-Histone H3 (Lys18) - 3171003 3171003
      ChIPAb+ Acetyl-Histone H3 (Lys18) - 3439954 3439954
      ChIPAb+ Acetyl-Histone H3 (Lys18) - 3714505 3714505
      ChIPAb+ Acetyl-Histone H3 (Lys18) -2547219 2547219
      ChIPAb+ Acetyl-Histone H3 (Lys18) -2744743 2744743
      ChIPAb+ Acetyl-Histone H3 (Lys18) -2800013 2800013
      ChIPAb+ Acetyl-Histone H3 (Lys18) -2856186 2856186
      ChIPAb+ Acetyl-Histone H3(Lys18) - NRG1849631 NRG1849631

      Brochure

      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