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

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
  • 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, all Mammals
  • 免疫原

    Synthetic peptide corresponding to Human Histone H3 aa 100 to the C-terminus (C terminal).
    Database link: P68431
    (Peptide available as ab12149)

  • ポジティブ・コントロール
    • This antibody gave a positive result in the following lysates: HeLa Whole Cell, Calf Thymus Histone and NIH3T3 (Mouse Embryonic Fibroblast) Whole Cell.

製品の特性

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 5 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
    • 別名
      • H3 histone family, member A antibody
      • H3/A antibody
      • H31_HUMAN antibody
      • H3FA antibody
      • Hist1h3a antibody
      • HIST1H3B antibody
      • HIST1H3C antibody
      • HIST1H3D antibody
      • HIST1H3E antibody
      • HIST1H3F antibody
      • HIST1H3G antibody
      • HIST1H3H antibody
      • HIST1H3I antibody
      • HIST1H3J antibody
      • histone 1, H3a antibody
      • Histone cluster 1, H3a 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

    Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade 画像

    • 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
      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.

    • 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.    

    • 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

    • 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.

    • 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
      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
    • 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, 5µg/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.

    Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) 使用論文

    This product has been referenced in:
    • Hernando H  et al. EZH2 Inhibition Blocks Multiple Myeloma Cell Growth through Upregulation of Epithelial Tumor Suppressor Genes. Mol Cancer Ther 15:287-98 (2016). Read more (PubMed: 26590165) »
    • Hacker KE  et al. Structure/Function Analysis of Recurrent Mutations in SETD2 Protein Reveals a Critical and Conserved Role for a SET Domain Residue in Maintaining Protein Stability and Histone H3 Lys-36 Trimethylation. J Biol Chem 291:21283-21295 (2016). Read more (PubMed: 27528607) »

    See all 20 Publications for this product

    Product Wall

    Abcam has not validated the combination of species/application used in this Abreview.
    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
    Username

    Mr. Andrew Lerner

    Verified customer

    投稿 Sep 25 2015

    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 (http://www.abcam.com/index....

    Read More

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

    Abcam has not validated the combination of species/application used in this Abreview.
    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
    Username

    Abcam user community

    Verified customer

    投稿 Apr 25 2011

    Abcam guarantees this product to work in the species/application used in this Abreview.
    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
    Username

    Abcam user community

    Verified customer

    投稿 Jan 10 2011

    Abcam has not validated the combination of species/application used in this Abreview.
    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
    Username

    Abcam user community

    Verified customer

    投稿 Jun 17 2010

    Unfortunately, we are not able to release the immunogen sequence for this particular antibody as it is proprietary information. We aim to provide as much information as possible to our customers, so I am sorry that this has not been possible on thi...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application ChIP
    Sample Rat Cell lysate - whole cell (Rat normal liver cells fresh isolated)
    Specification Rat normal liver cells fresh isolated
    Type Cross-linking (X-ChIP)
    Duration of cross-linking step: 15 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde
    Detection step Real-time PCR
    Negative control Normal IgG
    Username

    Abcam user community

    Verified customer

    投稿 Sep 25 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application ChIP
    Sample Mouse Cell lysate - whole cell (Liver, Cultured Hepatoma)
    Specification Liver, Cultured Hepatoma
    Type Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde
    Detection step Real-time PCR
    Positive control Anti RNA polymerase II
    Negative control Normal mouse IgG
    Username

    Abcam user community

    Verified customer

    投稿 Jul 30 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (lymphoma and liver cancer cells tested)
    Loading amount 2 µg
    Specification lymphoma and liver cancer cells tested
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: rt°C
    Username

    Abcam user community

    Verified customer

    投稿 Apr 21 2008

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