Anti-Histone H3 (phospho S10)抗体- ChIP Grade (ab5176)

概述

  • 产品名称Anti-Histone H3 (phospho S10)抗体- ChIP Grade
    参阅全部 Histone H3 一抗
  • 描述
    兔多克隆抗体to Histone H3 (phospho S10) - ChIP Grade
  • 特异性This antibody is specific for phosho S10 of histone H3. We believe that it does not recognise the non-modified histone - no blocking is seen with the non-phospho peptide (see Western blot image).
  • 经测试应用适用于: ICC/IF, WB, IHC-P, IHC - Wholemount, IHC-Fr, Flow Cyt, ChIP, IHC (PFA fixed), IHC-FoFrmore details
  • 种属反应性
    与反应: Mouse, Rat, Hamster, Cow, Human, Xenopus laevis, Drosophila melanogaster, Indian Muntjac, Zebrafish, African Green Monkey, Apteronotus leptorhynchus, Cyanidioschyzon merolae, Hydractinia echinata
    预测可用于: Saccharomyces cerevisiae, Caenorhabditis elegans, Schizosaccharomyces pombe, Neurospora crassa
  • 免疫原

    Synthetic peptide corresponding to Human Histone H3 aa 1-100 (phospho S10) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P68431
    (Peptide available as ab11477)

  • 阳性对照
    • Colcemid-treated HeLa histone Preparation; Hela whole cell extract IHC-P: Human normal kidney FFPE tissue sections.

性能

  • 形式Liquid
  • 存放说明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.
  • 存储溶液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...
  • 纯度Immunogen affinity purified
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab5176 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
ICC/IF Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human Histone H3 (phospho S10) peptide (ab11477).
IHC-P Use a concentration of 5 µg/ml.
IHC - Wholemount 1/250. PubMed: 20144789
IHC-Fr 1/200. (see Abreview)
Flow Cyt 1/1000.

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

ChIP Use at an assay dependent concentration. PubMed: 20089855
IHC (PFA fixed) Use at an assay dependent concentration. PubMed: 25359725
IHC-FoFr Use at an assay dependent concentration. PubMed: 19622634

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
    see all
  • 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

Anti-Histone H3 (phospho S10) antibody - ChIP Grade 图像

  • All lanes : Anti-Histone H3 (phospho S10) antibody - ChIP Grade (ab5176) at 1/5000 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
    HRP goat anti-rabbit (H+L) at 1/20000 dilution

    Predicted band size : 17 kDa


    Exposure time : 1 second
  • IHC image of ab5176 staining Histone H3 (phospho S10) in human kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab5176, 5µg/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. No primary antibody was used in the negative control (shown on the inset).

    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.

  • This image was kindly supplied by Prof Bryan Turner, University of Birmingham. Female Human Lymphoblastoid cells were incubated with ab5176.
  • SKN cells stained with ab5176 (green) at a dilution of 1/100. The cells were fixed in paraformaldehyde for 10 minutes prior to incubation with ab5176. The DNA is stained with DAPI (blue). 100x magnification.
  • All lanes : Anti-Histone H3 (phospho S10) antibody - ChIP Grade (ab5176) at 1 µg/ml

    Lane 1 : untreated histones
    Lane 2 : colcemid treated histones
    Lane 3 : untreated histones with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 4 : colcemid treated histones with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 5 : untreated histones with Human Histone H3 (unmodified ) peptide (ab2903) at 1 µg/ml
    Lane 6 : colcemid treated histones with Human Histone H3 (unmodified ) peptide (ab2903) at 1 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 17 kDa


    Exposure time : 10 seconds
  • ab5176 at a 1/100 dilution staining Drosophila melanogaster (wild type) polytene chromosomes by ICC/IF. The cells were formaldehyde fixed and blocked with 1% BSA prior to incubation with the antibody for 12 hours. Bound antibody was detected using a Cy3 conjugated goat anti-rabbit antibody.

    See Abreview

  • ab5176 (1/1000) staining a population of human HeLa cells positive for Histone H3 (phospho S10). Cells were trypsizined, pelleted and fixed in ice cold Ethanol. Debris was eliminated and FL2-A/FL2-W was used to eliminate clumping cells. For further experimental details please refer to abreview folder.

    See Abreview

  • IHC-Wholemount image of Histone H3 (phospho S10) ab5176 on zebrafish jaw. 5 day old zebrafish were permeabilized with proteinase K (10ug/ml, 50min, 37degrees), prior to staining. Staining visible in mitotic cells. Image shows 2 cells labelled with H3 phospho S10 antibody (green), nuclei are labelled with DAPI (blue).

    See Abreview


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 17 kDa

    Lysates: Lane 1,3,5: 0.5µg/lane untreated histone Lane 2,4,6: 0.5µg/lane colcemid treated histone - treated Primary antibody: Lane 1-2 : ab5176 (1?g/ml) Lane 3-4 :ab5176 (1?g/ml) + phospho peptide (ab5176) 1?g/ml Lane 5-6 :ab5176 (1?g/ml) + non-phospho peptide ( ab2903 batch 3) 1?g/ml

Anti-Histone H3 (phospho S10) antibody - ChIP Grade (ab5176)参考文献

This product has been referenced in:
  • Oblinger JL  et al. Components of the eIF4F complex are potential therapeutic targets for malignant peripheral nerve sheath tumors and vestibular schwannomas. Neuro Oncol 18:1265-77 (2016). Read more (PubMed: 26951381) »
  • Al-Lamki RS  et al. Tumor necrosis factor receptor 2-signaling in CD133-expressing cells in renal clear cell carcinoma. Oncotarget 7:24111-24 (2016). Human . Read more (PubMed: 26992212) »

See all 49 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Cardiomyocytes)
Gel Running Conditions Reduced Denaturing
Loading amount 25 µg
Specification Cardiomyocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Nov 14 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 500000 cells
Specification Canine Kidney Epithelial MDCK cells
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
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提交于 Sep 27 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Hamster Cell lysate - whole cell (BHK-21 (Kidney Fibroblasts))
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 50000 cells
Specification BHK-21 (Kidney Fibroblasts)
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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提交于 Jul 22 2016

Application Western blot
Sample Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 500000 cells
Treatment treated with IFNbeta for 24 hours
Specification mouse embryonic fibroblasts
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
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提交于 Feb 11 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (human Foreskin Fibroblasts)
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 100000 cells
Specification human Foreskin Fibroblasts
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
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提交于 Jan 14 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Cardiomyocytes)
Specification Cardiomyocytes
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
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提交于 Oct 12 2015

Application Western blot
Sample Human Cell lysate - whole cell (Hep3B)
Gel Running Conditions Non-reduced Denaturing (15%)
Loading amount 30 µg
Specification Hep3B
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
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提交于 Jun 19 2015

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Brain)
Permeabilization Yes - Triton x-100, 0.01%
Specification Brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
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提交于 Jun 17 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step 3%BSA+0.1% Triton X100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
Sample Mouse Cell (primary hepatoctyes)
Specification primary hepatoctyes
Permeabilization No
Fixative Formaldehyde
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提交于 Dec 19 2014

Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Mouse Cultured Cells (Cardiomyocytes)
Specification Cardiomyocytes
Permeabilization Yes - Triton x-100, 0.01%
Fixative Formaldehyde
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Dr. Shyam Sundar Nandi

Verified customer

提交于 Nov 20 2014

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