重组
RabMAb

Anti-Histone H3 (phospho S10 + T11)抗体[E173] (ab32107)

概述

  • 产品名称
    Anti-Histone H3 (phospho S10 + T11)抗体[E173]
    参阅全部 Histone H3 一抗
  • 描述
    兔单克隆抗体[E173] to Histone H3 (phospho S10 + T11)
  • 特异性
    This antibody detects Histone H3 phosphorylated on both Serine 10 and Threonine 11. However, the antibody shows higher affinity for phosphorylated Serine 10 than for phosphorylated Threonine 11. This was validated by ELISA, Dot Blot and WB peptide blocking experiments.
  • 经测试应用
    适用于: ELISA, Dot blot, ICC/IF, WB, IHC-P, IPmore details
    不适用于: Flow Cyt
  • 种属反应性
    与反应: Mouse, Rat, Human, Drosophila melanogaster
    预测可用于: a wide range of other species
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Histone H3 aa 1-100 (phospho S10).
    Database link: Q16695

  • 阳性对照
    • WB: HeLa cell lysate treated with Calyculin A. IHC-P: Lymphoma tissue. ICC/IF: HeLa cells
  • 常规说明

    A trial size is available for this product.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Store at 4°C prior to reconstitution. Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • 存储溶液
    PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • 克隆
    单克隆
  • 克隆编号
    E173
  • 同种型
    IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab32107 in the following tested applications.

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

应用 Ab评论 说明
ELISA Use at an assay dependent concentration.
Dot blot 1/1000.
ICC/IF 1/250 - 1/1000.
WB 1/2500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
IHC-P Use at an assay dependent concentration.
IP 1/40.
  • 应用说明
    Is unsuitable for Flow Cyt.
  • 靶标

    • 功能
      Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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.
    • 序列相似性
      Belongs to the histone H3 family.
    • 发展阶段
      Expressed throughout the cell cycle independently of DNA synthesis.
    • 翻译后修饰
      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.
      Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5 (H3K4me), Lys-37 and Lys-80. 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), which are linked to gene repression, are underrepresented. 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. Phosphorylation on Ser-32 (H3S31ph) is specific to regions bordering centromeres in metaphase chromosomes.
      Ubiquitinated. Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination.
    • 细胞定位
      Nucleus. Chromosome.
    • Information by UniProt
    • 数据库链接
    • 别名
      • H3 3 like sequence MH921 antibody
      • H3 3A antibody
      • H3 a antibody
      • H3 b antibody
      • H3 c antibody
      • H3 d antibody
      • H3 f antibody
      • H3 h antibody
      • H3 histone family member E pseudogene antibody
      • H3 i antibody
      • H3 j antibody
      • H3 k antibody
      • H3 l antibody
      • H33_HUMAN antibody
      • H3F3 antibody
      • H3f3b antibody
      • Histone H3 3 pseudogene antibody
      • Histone H3.3 antibody
      see all

    图片

    • All lanes : Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) at 1/10000 dilution

      Lane 1 : Untreated HeLa cell lysate
      Lane 2 : HeLa cell lysate treated with calyculin A
      Lane 3 : HeLa cell lysate treated with calyculin A and alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


      Exposure time : 1 second

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling Histone H3 (phospho S10 + T11) with ab32107 at a dilution of 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200, 2.5 μg/mL) - Microtubule Marker (Alexa Fluor® 594). DAPI (blue) was used as a nuclear counterstain.

      Confocal image showing nuclear staining on mitotic HeLa cell.

    • All lanes : Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) at 1/1000 dilution

      Lane 1 : HeLa whole cell lysate -treated with Calyculin A with Histone H3 phospho S10 peptide
      Lane 2 : HeLa whole cell lysate -treated with Calyculin A with Histone H3 phospho T11 peptide
      Lane 3 : HeLa whole cell lysate -treated with Calyculin A with unmodified Histone H3 peptide
      Lane 4 : HeLa whole cell lysate -treated with Calyculin A with no peptide

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Predicted band size : 15 kDa
      Observed band size : 15 kDa


      Exposure time : 1 second

      Blocking/Diluting buffer 5% NFDM/TBST

    • Dot blot analysis of Histone H3 phospho S10 peptide (Lane 1), Histone H3 phospho T11 peptide (Lane 2) and Histone H3 unmodified peptide (Lane 3) labelling Histone H3 (phospho S10 + T11) with ab32107 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) was used as the secondary antibody at a dilution of 1/10 000.

      Blocking and diluting buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes. 

    • Ab32107, at a 1/50 dilution, staining Histone H3 (phospho S10 + T11) in paraffin embedded lymphoma tissue by Immunohistochemistry.

    • ab32107 (1/1000) staining Histone H3 (phospho S10 + T11) in paraformaldehyde-fixed, DAPI counterstained HeLa cells. Secondary antibody: Goat anti-Rabbit IgG conjugated to Cy3® (1/200). Please refer to abreview for further details.

      See Abreview

    • Direct ELISA antibody dose-response curve using ab32107 (0-1000 ng/mL).

      Peptides - Histone H3 unmodified peptide, Histone H3 phospho S10 peptide, Histone H3 phospho T11 peptide (100 ng/mL).

      Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500).

    • Direct ELISA antibody dose-response curve using ab32107 (0-1000 ng/mL).

      Peptides - Histone H3 unmodified peptide, Histone H3 phospho S10 peptide, Histone H3 phospho T11 peptide (10 ng/mL).

      Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500).

    文献

    This product has been referenced in:
    • Rabinowicz N  et al. Targeting the centriolar replication factor STIL synergizes with DNA damaging agents for treatment of ovarian cancer. Oncotarget 8:27380-27392 (2017). Read more (PubMed: 28423708) »
    • Zhang YL  et al. Development and characterization of a novel long-acting recombinant follicle stimulating hormone agonist by fusing Fc to an FSH-ß subunit. Hum Reprod 31:169-82 (2016). Read more (PubMed: 26621853) »

    See all 20 Publications for this product

    客户评价及客户问答

    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Fruit fly (Drosophila melanogaster) Tissue lysate - whole (0-4hr embryos)
    Specification
    0-4hr embryos
    Treatment
    control shRNA and wee shRNA embryos
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Username

    Dr. Richelle Sopko

    Verified customer

    提交于 Dec 04 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Username

    Dr. Kirk Mcmanus

    Verified customer

    提交于 Feb 20 2007

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

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