Anti-Histone H3 抗体 [mAbcam 10799] - ChIP Grade (ab10799)

Reviews (11)Q&A (3)References (70)

製品の概要

  • 製品名

    Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade
    Histone H3 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [mAbcam 10799] to Histone H3 - ChIP Grade

  • 由来種

    Mouse

  • アプリケーション

    適用あり: ChIP, WB, IHC-P, IPmore details
    適用なし: ICC/IF

  • 種交差性

    交差種: Mouse, Rat, Human, Drosophila melanogaster, Monkey
    交差が予測される動物種: a wide range of other species, Mammals

  • 免疫原

    Synthetic peptide within Human Histone H3 aa 100 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P68431
    (Peptide available as ab12149)

  • ポジティブ･コントロール

    • WB: HeLa, Calf Thymus Histone and NIH3T3 (Mouse Embryonic Fibroblast) Whole Cell Lysates; IP: HeLa whole cell extract; IHC-P: FFPE human breast adenocarcinoma.

  • 特記事項

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...

  • 精製度

    IgG fraction

  • ポリ/モノ

    モノクローナル

  • クローン名

    mAbcam 10799

  • ミエローマ

    Sp2/0-Ag14

  • アイソタイプ

    IgG3

  • 軽鎖の種類

    kappa

  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab10799 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
ChIP Use 2-4 µg for 25 µg of chromatin.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use a concentration of 1 µg/ml.
  • 追加情報
    Is unsuitable for ICC/IF.

    • ターゲット情報

    • 機能

      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.

    • 配列類似性

      Belongs to the histone H3 family.

    • 発生段階

      Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.

    • 翻訳後修飾

      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.

    • 細胞内局在

      Nucleus. Chromosome.
    • Information by UniProt

    • 参照データベース

      see all

    • 別名

      • H3 histone family member E pseudogene antibody
      • H3 histone family, member A antibody
      • H3/A antibody
      • H31_HUMAN antibody
      • H3F3 antibody
      • H3FA antibody
      • Hist1h3a antibody
      • HIST1H3B antibody
      • HIST1H3C antibody
      • HIST1H3D antibody
      • HIST1H3E antibody
      • HIST1H3F antibody
      • HIST1H3G antibody
      • HIST1H3H antibody
      • HIST1H3I antibody
      • HIST1H3J antibody
      • HIST3H3 antibody
      • histone 1, H3a antibody
      • Histone cluster 1, H3a antibody
      • Histone H3 3 pseudogene antibody
      • Histone H3.1 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

    画像

    • ChIP - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
      ChIP - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

      Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 2µg of ab10799 (blue), and 20µl 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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.    

    • Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
      Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
      All lanes : Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) at 5 µg/ml

      Lane 1 : Histone H1 Recombinant Protein
      Lane 2 : Histone H2A Recombinant Protein
      Lane 3 : Histone H2B Recombinant Protein
      Lane 4 : Histone H3.1 Recombinant Protein
      Lane 5 : Histone H4 Recombinant Protein

      Lysates/proteins at 0.1 µg per lane.

      Secondary
      All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 15 kDa


      Exposure time: 4 minutes


      We recommend using 3% milk as the blocking agent in Western Blot.

    • Immunoprecipitation - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
      Immunoprecipitation - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

      Histone H3 [mAbcam 10799] - ChIP Grade was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Mouse monoclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

      The antibody was incubated under agitation with Protein G beads for 10min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab10799.

      Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

      Band: 15kDa; [mAbcam 10799] Histone H3 - ChIP Grade

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

      ab10799 staining human kidney sections by IHC-P using EXPOSE IHC detection kit (ab80436). Formalin fixed paraffin embedded tissue sections were pre-treated using heat mediated antigen retrieval (using a pressure cooker) with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab10799, 5µg/ml, for 1 hour at room temperature. DAB was used as the chromogen and the section was counterstained with haematoxylin and mounted with DPX.

    • Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
      Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
      All lanes : Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
      Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
      Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

      Secondary
      All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Performed under reducing conditions.

      Predicted band size: 15 kDa
      Observed band size: 17 kDa
      why is the actual band size different from the predicted?


      Exposure time: 90 seconds
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

      IHC image of Histone H3 staining in human breast carcinoma FFPE section, performed on a 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 ab10799, 1µg/ml, for 8 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.

    参考文献

    This product has been referenced in:

    See all 70 Publications for this product

    ab10799 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    レビューと Q&A

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    レビューを送る 質問を送る

    1-10 of 14 Abreviews or Q&A

    Western blot abreview for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (primary myoblast)
    Gel Running Conditions
    Reduced Denaturing (4-20)
    Loading amount
    10 µg
    Specification
    primary myoblast
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jul 18 2019

    IHC - Wholemount abreview for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Xenopus laevis Embryo (Whole-embryo IHC, early stage (stage 21))
    Specification
    Whole-embryo IHC, early stage (stage 21)
    Read More

    Dr. Clotilde Cadart

    Verified customer

    投稿 Mar 26 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

     Excellent
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Whole brain sections)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Permeabilization
    Yes - 0.5% Tween-20
    Specification
    Whole brain sections
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Oct 26 2018

    Western blot abreview for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

     Excellent
    Abreviews
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Whole brain tissue lysate)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (3-8% Bis-Tris)
    Loading amount
    10 µg
    Specification
    Whole brain tissue lysate
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 29 2017

    Western blot abreview for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Saccharomyces cerevisiae Cell lysate - whole cell (yeast)
    Gel Running Conditions
    Non-reduced Denaturing (15)
    Loading amount
    2e+006 cells
    Specification
    yeast
    Read More

    Mr. Andrew Lerner

    Verified customer

    投稿 Sep 25 2015

    Question


    Mon équipe a tenté de marquer ces anticorps avec nos Dye et nous avons eu un soucis sur l'anticorps 10799 qui ne s'est quasiment pas marqué. Pourriez vous me confirmer qu'il n'y a absolument pas de BSA dans le tampon de cet anticorps?

    Read More
    Answer

    Je peux vous confirmer que cet anticorps ne contient pas de BSA. Cependant, il contient 0.4M d’arginine qui pourrait interférer avec la conjugaison du fluorochrome. Comme expliqué sur notre site internet (https://www.abcam.com/index.html?pageconfig=resource&rid=13156), nous recommandons pour nos kits de conjugaison « Easylink » d’éviter les nucléophiles tels que des acides aminés dans les tampons. Ces acides aminés pourraient désactiver le produit chimique qui lie de manière covalente le fluorochrome à l'anticorps.

    Read More

    Question

    This is a clinical diagnostics project, and all antibodies we will use for that need to be CE certified for diagnostic purposes. Since I did not find any information on this on your website, I wanted to ask you if you have any CE certified antibodies directed against these (human) proteins:

    Read More
    Answer

    All of our products are for research purposes only and are not certified for clinical research applications.

    Read More

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

     Poor
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (sputum)
    Specification
    sputum
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.005% saponin
    Blocking step
    Block Ace(Dainipon Sumitomo Seiyaku) as blocking agent for 10 minute(s) · Concentration: 20% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 25 2011

    Western blot abreview for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (PC-3 (Metastatic Prostate Cancer Cell Line))
    Loading amount
    15 µg
    Specification
    PC-3 (Metastatic Prostate Cancer Cell Line)
    Gel Running Conditions
    Reduced Denaturing (10)
    Blocking step
    Aqua Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 10 2011

    Western blot abreview for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade

     Inconclusive
    Abreviews
    Application
    Western blot
    Sample
    Saccharomyces cerevisiae Cell lysate - whole cell (TCA extraction protocol)
    Loading amount
    30 µg
    Specification
    TCA extraction protocol
    Gel Running Conditions
    Reduced Denaturing (12% Tris-HCl)
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jun 17 2010

    1-10 of 14 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com