Anti-Histone H3 (phospho S10)抗体[mAbcam 14955] - ChIP Grade (ab14955)

概述

  • 产品名称Anti-Histone H3 (phospho S10)抗体[mAbcam 14955] - ChIP Grade
    参阅全部 Histone H3 一抗
  • ��述
    小鼠单克隆抗体[mAbcam 14955] to Histone H3 (phospho S10) - ChIP Grade
  • 特异性ab14955 recognises phospho S10 on Histone H3, even when K9 is tri methylated. It recognises a phospho S28 peptide by ELISA, but is not blocked by phospho S28 in WB.
  • 经测试应用适用于: IHC-P, ICC/IF, ChIP, IP, IHC-Fr, Flow Cyt, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Xenopus laevis, Arabidopsis thaliana, Drosophila melanogaster, Indian Muntjac, African Green Monkey
    预测可用于: Chicken, Saccharomyces cerevisiae, Caenorhabditis elegans, Schizosaccharomyces pombe, Zebrafish, all Mammals, Tobacco, Chlamydomonas reinhardtii, Aspergillus nidulans, Neurospora crassa
  • 免疫原

    Phospho S10 specific clone produced using a synthetic peptide derived from residues 1 - 100 of Human Histone H3, phosphorylated at S10 and tri methylated at K9.

    Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising tri methyl K9 and phospho S10 dimodified peptide. Clones were also positively screened against both tri methyl K9 and phospho S10 peptides. Clones were negatively screened against the unmodified version of the peptides. This clone binds to the tri methyl K9 and phospho S10 dimodified peptide and to the phospho S10 peptide, but not to the tri methyl K9 peptide or to equivalent unmodified Histone H3 peptide.

  • 阳性对照
    • IHC-P: Human normal kidney FFPE tissue sections.
  • 常规说明

    Alternative versions available:

    Anti-Histone H3 antibody (phospho S10) (Alexa Fluor® 488) [mAbcam 14955] (ab197502)

    Anti-Histone H3 antibody (phospho S10) (Alexa Fluor® 647) [mAbcam 14955] (ab196698)

性能

应用

Our Abpromise guarantee covers the use of ab14955 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
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration. PubMed: 20864037See Abreview submitted on 5 January 2007.
IP Use at 80 µg/mg of lysate.
IHC-Fr Use at an assay dependent concentration. PubMed: 21734301
Flow Cyt 1/1000.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).

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 [mAbcam 14955] - ChIP Grade 图像

  • All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] - ChIP Grade (ab14955) at 1/2000 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 : 15 kDa


    Exposure time : 1 second
  • Indian muntjac and HeLa cells were fixed in paraformaldehyde and labelled with ab14955 (1/2000 dilution) for 30 minutes. The above image contains an interphase and a prophase Indian muntjac cells immunofluorescently labelled with anti-PhosS10 (green) and counterstained with DAPI (red).

  • All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] - ChIP Grade (ab14955) at 1 µg/ml

    Lane 1 : Control HeLa Histone Prep 0.5ug
    Lane 2 : Colcemid treated HeLa Histone prep 0.5ug
    Lane 3 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
    Lane 4 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
    Lane 5 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
    Lane 6 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
    Lane 7 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 8 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 9 : Control HeLa Histone Prep 0.5ug with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
    Lane 10 : Colcemid treated HeLa Histone prep 0.5ug with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml

    Secondary
    Rabbit polyclonal to Mouse IgG H&L (HRP) at 1/5000 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?)
  • IHC image of ab14955 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 ab14955, 1µg/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.

  • All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] - ChIP Grade (ab14955) at 1/1000 dilution

    Lane 1 : Wild type 0-4 hour old fruit fly embryo lysate
    Lane 2 : 0-4 hour old fruit fly embryo lysate expressing wee RNAi

    Secondary
    Anti-mouse IgG, peroxidase-linked at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa


    Exposure time : 5 seconds

    Image courtesy of Richelle Sopko, Harvard University, U.S.A

    Blocking: 10% BSA

    wee shRNA embryos (lane 2) should display elevated phospho H3Ser10 levels relative to wild type

    See Abreview

  • All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] - ChIP Grade (ab14955) at 0.5 µg/ml

    Lane 1 : Control HeLa Histone prep
    Lane 2 : Colecemid treated HeLa Histone prep
    Lane 3 : Control HeLa Histone prep with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
    Lane 4 : Colecemid treated HeLa Histone prep with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
    Lane 5 : Control HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
    Lane 6 : Colecemid treated HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
    Lane 7 : Control HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 8 : Colecemid treated HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml

    Lysates/proteins at 5 µg per lane.

    Secondary
    Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

    Predicted band size : 15 kDa
    Observed band size : 20 kDa (why is the actual band size different from the predicted?)

    Lane 1 - 8 : Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 0.5 ug/ml

    Lane 1 : Control HeLa Histone prep at 5 ug
    Lane 2 : Colcemid treated HeLa Histone prep at 5 ug
    Lane 3 : Control HeLa Histone prep at 5 ug with Histone H3 peptide - unmodified (ab7228) at 1 ug/ml
    Lane 4 : Colcemid treated HeLa Histone prep at 5 ug with Histone H3 peptide - unmodified (ab7228) at 1 ug/ml
    Lane 5 : Control HeLa Histone prep at 5 ug with Histone H3 peptide - phospho S28 (ab14793) at 1 ug/ml
    Lane 6 : Colcemid treated HeLa Histone prep at 5 ug with Histone H3 peptide - phospho S28 (ab14793) at 1 ug/ml
    Lane 7 : Control HeLa Histone prep at 5 ug with Histone H3 peptide - phospho S10 (ab11477) at 1 ug/ml

  • ab14955 (1/1000) stainning a population of Human HeLa cells positive for Histone H3 (phospho S10). Cells were trypsizined, pelleted and fixed in ice cold ethanol. Cellular debris was eliminated and the FL2-A/FL2-W was used to eliminate clumping cells. For further experimental details please refer to abreview.

    See Abreview

  • By ELISA, ab14955 detects:
    the singly modified phospho S10 peptide and the dual modified phospho S10 and tri methyl K9 peptide (the orange and grey lines respectively),
    less strongly detects the phospho S28 peptide (purple line),
    does not detect the equivalent non-modified Histone H3 peptide for S10 or the singly modified tri methyl K9 peptide (the 2 lines at the bottom of the figure).

    The antibody recognises phospho S28 by ELISA (although a phospho S28 peptide does not block the antibody in Western blotting).

Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] - ChIP Grade (ab14955)参考文献

This product has been referenced in:
  • Staples CJ  et al. MRNIP/C5orf45 Interacts with the MRN Complex and Contributes to the DNA Damage Response. Cell Rep 16:2565-75 (2016). Read more (PubMed: 27568553) »
  • 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 35 Publications for this product

Product Wall

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 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Arabidopsis thaliana Cell (flower buds)
Specification flower buds
Fixative Paraformaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
Username

Dr. Dmitri Demidov

Verified customer

提交于 Aug 30 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (H1299)
Loading amount 5 µg
Specification H1299
Treatment 100 ng/ml PMA for 2 h
Gel Running Conditions Reduced Denaturing (15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. yipeng du

Verified customer

提交于 Mar 01 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (E18 Mouse Brain Sections)
Specification E18 Mouse Brain Sections
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization Yes - 1% Triton in PBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Apr 20 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample African Green Monkey Cell (COS-7 (and Indian Muntjac [IM]))
Specification COS-7 (and Indian Muntjac [IM])
Fixation Ice Cold Ethanol
Permeabilization No
Gating Strategy Cellular debris was eliminated and the FL2-A/FL2-W was used to eliminate clumping cells
Username

Dr. Kirk McManus

Verified customer

提交于 Dec 10 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (10T1/2)
Specification 10T1/2
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk McManus

Verified customer

提交于 Dec 10 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (HeLa)
Specification HeLa
Fixation Ice cold ethanol
Permeabilization No
Gating Strategy Cellular debris was eliminated and the FL2-A/FL2-W was used to eliminate clumping cells
Username

Dr. Kirk McManus

Verified customer

提交于 Nov 10 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - nuclear (Nuclear extract from Rat2 cells)
Loading amount 15 µg
Specification Nuclear extract from Rat2 cells
Gel Running Conditions Non-reduced Denaturing (4-20% tricine gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Oct 10 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (embryonic brain section)
Specification embryonic brain section
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization Yes - 0.1% Triton-X
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jun 17 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Caenorhabditis elegans Cell lysate - whole cell (Embryo)
Total protein in input 250 µg
Specification Embryo
Immuno-precipitation step Protein G
Username

Dr. Caroline Thivierge

Verified customer

提交于 Apr 15 2008

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