Anti-Histone H4 (mono methyl K20) 抗体 - ChIP Grade (ab9051)

製品の概要

  • 製品名
    Anti-Histone H4 (mono methyl K20) antibody - ChIP Grade
    Histone H4 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Histone H4 (mono methyl K20) - ChIP Grade
  • アプリケーション
    適用あり: IHC-P, IHC-Fr, WB, ICC/IF, Flow Cyt, CHIPseq, ChIPmore details
  • 種交差性
    交差種: Mouse, Cow, Human, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe
    交差が予測される動物種: Mammal
  • 免疫原

    Synthetic peptide within Human Histone H4 aa 1-100 (mono methyl K20) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab17043)

  • ポジティブ・コントロール
    • WB: Calf thymus histone preparation and HeLa whole cell extract. IHC-P: Human normal skin tissue.
  • 特記事項

    Interphase: The cells within the culture show a considerable variability in the intensity of staining with the antibody. The relationship between trimethylation levels and cell cycle have not yet been determined but may be a contributor to the amount of methylation detected in each cell. Heterochromatic regions of the interphase nucleus are the primary sites of trimethylation observed by indirect immunofluroescence. Mitosis: Discrete chromosomal regions are labelled intensely, with lower level fluorescence throughout the remainder of the chromosome arms Data from IF performed by Kirk McManus in the lab of Michael Hendzel as part of the antibody characterisation at cellnucleus.com

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped 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.
  • バッファー
    pH: 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...
  • 精製度
    Immunogen affinity purified
  • 一次抗体 備考
    Interphase: The cells within the culture show a considerable variability in the intensity of staining with the antibody. The relationship between trimethylation levels and cell cycle have not yet been determined but may be a contributor to the amount of methylation detected in each cell. Heterochromatic regions of the interphase nucleus are the primary sites of trimethylation observed by indirect immunofluroescence. Mitosis: Discrete chromosomal regions are labelled intensely, with lower level fluorescence throughout the remainder of the chromosome arms Data from IF performed by Kirk McManus in the lab of Michael Hendzel as part of the antibody characterisation at cellnucleus.com
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab9051 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-P Use a concentration of 0.2 µg/ml.
IHC-Fr Use at an assay dependent concentration.
WB 1/1000. Can be blocked with Human Histone H4 (mono methyl K20) peptide (ab17043).
ICC/IF Use at an assay dependent concentration. Cautionary note: At low dilutions, the antibody may cross-react with microtubules. This can be minimized by increasing the dilution of the antibody when used in immunofluroescence.
Flow Cyt 1/200.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

CHIPseq Use at an assay dependent concentration. PubMed: 22196736
ChIP Use 2-25 µg for 25 µg of chromatin.

ターゲット情報

Anti-Histone H4 (mono methyl K20) antibody - ChIP Grade 画像

  • 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  ab9051 (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.

  • All lanes : Anti-Histone H4 (mono methyl K20) antibody - ChIP Grade (ab9051) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 peptide (ab14963) at 0.5 µg/ml
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 (mono methyl K20) peptide (ab17043) at 0.5 µg/ml
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Histone H4 peptide (17-24) - di methyl K20 at 0.5 µg/ml
    Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H4 (tri methyl K20) peptide (ab17567) at 0.5 µg/ml
    Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
    Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
    Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml

    Lysates/proteins at 0.5 µg per lane.

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

    Performed under reducing conditions.

    Observed band size : 13 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 seconds
  • Rabbit polyclonal to Histone H4 mono methyl K20 (1/250).

    HeLa cells cultured on glass coverslips were fixed with 4% paraformaldehyde and then stained with ab9051 (green). Total chromatin was visualized using DAPI staining (red).

    This image is part of the antibody characterisation at www.cellnucleus.com

  • IHC image of Histone H4 (mono methyl K20) 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 ab9051, 0.2µg/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.

    Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • IHC - Wholemount of Caenorhabditis elegans embryo labelling Histone H4 (mono methyl K20) with ab9051. Sample was incubated with primary antibody (1/400 in TBS + 0.1% Triton X-100 + 3% Goat serum) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody. Left - Histone H4 (mono methyl K20), Middle - DAPI, Right - merge.

    See Abreview

  • Rabbit polyclonal to Histone H4 mono methyl K20 at 1/1000 on histone prep (Sigma).

    Peptide competition (shown above lanes) at 1ug/ml.

    This peptide blocking study shows that ab9051 is specific for Histone H4 (mono methyl K20). The activity of the antibody is specifically blocked by the addition of the H4 mono methyl peptide (ab17043).

  • Rabbit polyclonal to Histone H4 mono methyl K20 (1/250).

    HeLa cells cultured on glass coverslips were fixed with 4% paraformaldehyde and then stained with ab9051 (green). Total chromatin was visualized using DAPI staining (red).

    Left: merge; center: DAPI; right: Mono K20

    This image is part of the antibody characterisation at www.cellnucleus.com

  • Rabbit polyclonal to Histone H4 mono methyl K20 (1/250).

    10T1 cells cultured on glass coverslips were fixed with 4% paraformaldehyde and then stained with ab9051.

    Top left: metaphase cell. Mono methyl K20 (green); DAPI (red)
    Top right: interphase cell. Mono methyl K20 (green); DAPI (red)
    Lower left: interphase cell. Mono methyl K20 (green); DAPI (blue), TBP (red)
    Lower right: interphase cell. Mono methyl K20 (green); DAPI (red)

    This image is part of the antibody characterisation at www.cellnucleus.com

Anti-Histone H4 (mono methyl K20) antibody - ChIP Grade (ab9051) 使用論文

This product has been referenced in:
  • Shao P  et al. Histone demethylase PHF8 promotes epithelial to mesenchymal transition and breast tumorigenesis. Nucleic Acids Res 45:1687-1702 (2017). ChIP ; Human . Read more (PubMed: 27899639) »
  • Asensio-Juan E  et al. The histone demethylase PHF8 is a molecular safeguard of the IFN? response. Nucleic Acids Res 45:3800-3811 (2017). ChIP ; Human . Read more (PubMed: 28100697) »

See all 75 Publications for this product

Product Wall

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Na-citrate pH6
Sample
Mouse Tissue sections (Liver, P5)
Specification
Liver, P5
Permeabilization
Yes - Triton 0,05%
Fixative
Paraformaldehyde
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投稿 Oct 07 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse embryonic fibroblasts)
Specification
Mouse embryonic fibroblasts
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 23°C
Fixative
Paraformaldehyde
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投稿 Nov 28 2013

Application
Western blot
Loading amount
8 µg
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Caenorhabditis elegans Tissue lysate - nuclear (C. elegans embryo)
Specification
C. elegans embryo
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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投稿 Nov 13 2013

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12.5%)
Sample
Mouse Cell lysate - nuclear (Mouse embryonic fibroblasts)
Specification
Mouse embryonic fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Nov 11 2013

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (4~15%)
Sample
Human Cell lysate - whole cell (Human HeLa cells)
Specification
Human 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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投稿 Nov 11 2013

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (15%)
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (Fruit fly embryo lysate)
Specification
Fruit fly embryo lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Nov 11 2013

Application
IHC - Wholemount
Sample
Caenorhabditis elegans Embryo (Whole embryo of C. elegans)
Specification
Whole embryo of C. elegans
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投稿 Oct 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Mouse Cell (fibroblast)
Specification
fibroblast
Permeabilization
Yes - 0.5% Triton
Fixative
Formaldehyde
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投稿 Sep 12 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (multiple myeloma cells)
Loading amount
50 µg
Specification
multiple myeloma cells
Gel Running Conditions
Reduced Denaturing (10-20% tris glycine)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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投稿 Oct 30 2012

Le laboratoire m'a indiqué que l'anticorps ab9051 est batch-testé en ChIP. Pourriez-vous m'envoyer le numéro de commande correspondant au lot xxxxxx? Je pourrai ainsi appliquer la garantie sur ce produit et vous envoyer gratuitement une unité d'...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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