Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191)

Overview

  • Product nameAnti-Histone H3 (acetyl K18) antibody - ChIP Grade
    See all Histone H3 primary antibodies
  • Description
    Rabbit polyclonal to Histone H3 (acetyl K18) - ChIP Grade
  • SpecificitySpecificity for acetyl K18 has been validated by blocking peptide/antibody incubation followed by immunostaining western blots.
  • Tested applicationsWB, PepArr, CHIPseq, IHC-P, ICC/IF, Flow Cyt, ChIP/Chip, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Saccharomyces cerevisiae, Arabidopsis thaliana, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Fungi
  • Immunogen

    Synthetic peptide derived from within residues 1 - 100 Histone H3, acetylated at K18.

    (Peptide available as ab24003.)

  • Positive control
    • This antibody gave a positive signal in HeLa Histone Preparation Nuclear Lysate. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal skin.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferpH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Clonality Polyclonal
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab1191 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 17 kDa.Can be blocked with Human Histone H3 (acetyl K18) peptide (ab24003).
PepArr Use a concentration of 0.2 - 0.02 µg/ml.
CHIPseq Use at an assay dependent concentration.
IHC-P 1/200.
ICC/IF Use a concentration of 1 µg/ml.
Flow Cyt Use at an assay dependent concentration.
ChIP/Chip Use at an assay dependent concentration.
ChIP Use 2µg for 106 cells.

Target

  • FunctionCore 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.
  • Sequence similaritiesBelongs to the histone H3 family.
  • Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • Post-translational
    modifications
    Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
    Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
    Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
    Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
    Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) 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 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
    Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.
  • Cellular localizationNucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H 3 antibody
    • H3 3 Like Sequence MH921 antibody
    • H3 3A antibody
    • H3 3A antibody
    • H3 antibody
    • H3 Histone antibody
    • H3 Histone Family Member E Pseudogene antibody
    • H3.4 antibody
    • H3/A antibody
    • H3/g antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3F3 antibody
    • H3FA antibody
    • H3FT antibody
    • H3t antibody
    • HIST1H3J antibody
    • HIST3H3 antibody
    • HIST3H3 antibody
    • Histone cluster 1, H3a antibody
    • Histone H3 3 Pseudogene antibody
    • Histone H3.1 antibody
    • Histone H3.3 antibody
    • Histone H3/a antibody
    • Histone H3/b antibody
    • Histone H3/c antibody
    • Histone H3/d antibody
    • Histone H3/f antibody
    • Histone H3/h antibody
    • Histone H3/i antibody
    • Histone H3/j antibody
    • Histone H3/k antibody
    • Histone H3/l antibody
    see all

Anti-Histone H3 (acetyl K18) antibody - ChIP Grade images

  • All batches of ab1191 are tested in Peptide Array against peptides to different Histone H3 and H4 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - acetyl K18 peptide (ab24003), indicating that this antibody specifically recognises the Histone H3 - acetyl K18 modification.

    ab17163 - Histone H3 - unmodified

    ab16635 - Histone H3 - acetyl K9

    ab15591 - Histone H3 - acetyl K14

    ab24003 - Histone H3 - acetyl K18

    ab48359 - Histone H3 - acetyl K23

    ab24404 - Histone H3 - acetyl K27

    ab41409 - Histone H3 - acetyl K36

    ab15662 - Histone H4 - acetyl K12

  • Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab1191 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

  • L929, 3T3 and HeLa cells fixed with 2% paraformaldehyde in PBS were incubated for 5 min in 0.2% Triton X-100 in PBS before a blocking step in 5% BSA, 0.1% Tween 20 in PBS (blocking buffer). Ac K18-H3 (ab1191) antibody was added diluted at 1/500 in blocking buffer for a 45 min incubation at room temperature. For visualization, fluorescein isothiocyanate (FITC) anti rabbit secondary antibody was used diluted at 1/800. DAPI (Sigma) in 0.1% Tween 20 in PBS was used in the final washing steps to stain DNA. Vectashield (Vector Laboratories Inc.) was used as mounting medium.
    The ac K18-H3 antibody revealed a granular staining throughout the nucleoplasm, excluding regions that show an intense DAPI staining (mostly pericentric heterochromatin in mice cells).
  • Lane 1 : Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191) at 1 µg/ml (Sample: Histone prep, 0.5 ug/ml )
    Lane 2 : Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191) at 1 µg/ml

    Lane 1 : As above
    Lane 2 : Human Histone H3 (acetyl K18) peptide (ab24003) at 1 µg/ml

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Predicted band size : 15.4 kDa
    Observed band size : 17 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab1191 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1191, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml.
  • All lanes : Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191) at 1 µg/ml

    Lane 1 : HeLa Histone Preparation Nuclear Lysate
    Lane 2 : HeLa Histone Preparation Nuclear Lysate with Histone H3 peptide - unmodified R17 at 0.5 µg/ml
    Lane 3 : HeLa Histone Preparation Nuclear Lysate with Human Histone H3 (acetyl K9) peptide (ab16635) at 0.5 µg/ml
    Lane 4 : HeLa Histone Preparation Nuclear Lysate with Histone H3 peptide - acetyl K14 at 0.5 µg/ml
    Lane 5 : HeLa Histone Preparation Nuclear Lysate with Human Histone H3 (acetyl K18) peptide (ab24003) at 0.5 µg/ml
    Lane 6 : HeLa Histone Preparation Nuclear Lysate with Human Histone H3 (acetyl K23) peptide (ab48359) at 0.5 µg/ml
    Lane 7 : HeLa Histone Preparation Nuclear Lysate with Human Histone H3 (acetyl K27) peptide (ab24404) at 0.5 µg/ml
    Lane 8 : HeLa Histone Preparation Nuclear Lysate with Histone H3 peptide - acetyl K36 at 0.5 µg/ml
    Lane 9 : HeLa Histone Preparation Nuclear Lysate with Histone H4 (acetyl K12) peptide (ab15662) at 0.5 µg/ml

    Lysates/proteins at 2.5 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15.4 kDa
    Observed band size : 17 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • IHC image of Histone H3 (acetyl K18) staining in Human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1191, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191)

This product has been referenced in:
  • Wang J  et al. Lysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation. PLoS Pathog 10:e1003830 (2014). WB . Read more (PubMed: 24391497) »
  • Mohan RD  et al. Loss of Drosophila Ataxin-7, a SAGA subunit, reduces H2B ubiquitination and leads to neural and retinal degeneration. Genes Dev 28:259-72 (2014). Fruit fly (Drosophila melanogaster) . Read more (PubMed: 24493646) »

See all 44 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - nuclear (whole tissue extract)
Specification whole tissue extract
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Aug 05 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Caenorhabditis elegans Tissue lysate - nuclear (whole tissue extract)
Specification whole tissue extract
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Aug 05 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Tissue lysate - nuclear (whole tissue extract)
Specification whole tissue extract
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Aug 05 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (whole tissue extract)
Specification whole tissue extract
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Aug 01 2014

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - eBioscience Permeabilization Buffer
Sample Human Cell (Differentiated Haematopoietic Stem Cells)
Specification Differentiated Haematopoietic Stem Cells
Gating Strategy Isotype negative control (white)
Preparation Cell harvesting/tissue preparation method: Typsin-EDTA Cell Dissociation
Sample buffer: PBS and 10% FBS
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Submitted Feb 18 2014

Application Western blot
Loading amount 15 µg
Gel Running Conditions Reduced Denaturing (15%)
Sample Human Cell lysate - whole cell (Huvec)
Specification Huvec
Treatment MS275
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 4°C
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Submitted Nov 05 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (Fruit flyer nuclear lysate)
Specification Fruit flyer nuclear lysate
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Submitted Oct 22 2013

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Human Cell lysate - whole cell (Hela cells)
Specification Hela cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Oct 14 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Caenorhabditis elegans Tissue lysate - nuclear (C. elegan whole tissue lysate)
Specification C. elegan whole tissue lysate
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 1°C
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Submitted Oct 14 2013

Western blot

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Abreviews
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Application Western blot
Loading amount 5 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (12% SDS gel)
Sample Fruit fly (Drosophila melanogaster) Tissue lysate - whole (Whole mosquito (Culex pipiens))
Specification Whole mosquito (Culex pipiens)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted May 30 2013

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