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Epigenetics and Nuclear Signaling Histones H3 Methyl + Phospho

Anti-Histone H3 (tri methyl K9,phospho S10)抗体(ab5819)

  • Datasheet
  • SDS
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)
  • Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)
  • Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)

Key features and details

  • Rabbit polyclonal to Histone H3 (tri methyl K9, phospho S10)
  • Suitable for: IHC-P, WB, ICC/IF
  • Reacts with: Mouse, Cow, Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-Histone H3 (tri methyl K9,phospho S10)抗体
    参阅全部 Histone H3 一抗
  • 描述

    兔多克隆抗体to Histone H3 (tri methyl K9,phospho S10)
  • 宿主

    Rabbit
  • 特异性

    From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help.

  • 经测试应用

    适用于: IHC-P, WB, ICC/IFmore details
  • 种属反应性

    与反应: Mouse, Cow, Human
  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: Calf Thymus Histone, HeLa Nuclear, NIH3T3 Nuclear. ICC/IF: SKN cells, HeLa cells. IHC-P: Human breast carcinoma.
  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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
    Constituents: 98.98% PBS, 1% BSA

    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

    • Epigenetics and Nuclear Signaling
    • Histones
    • H3
    • Methyl + Phospho

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Control Peptide

    • Human Histone H3 (tri methyl K9) peptide (ab1773)
  • Corresponding Unmodified Peptide

    • Human Histone H3 (unmodified) peptide (ab7228)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human Histone H3 protein (Active) (ab198757)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab5819于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB
1/500 - 1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 17 kDa).
ICC/IF (1)
Use at an assay dependent concentration.
说明
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB
1/500 - 1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 17 kDa).
ICC/IF
Use at an assay dependent concentration.

靶标

  • 功能

    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.
  • 序列相似性

    Belongs to the histone H3 family.
  • 发展阶段

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

    Nucleus. Chromosome.
  • Target information above from: UniProt accession P68431 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 8350 Human
    • Entrez Gene: 8351 Human
    • Entrez Gene: 8352 Human
    • Entrez Gene: 8353 Human
    • Entrez Gene: 8354 Human
    • Entrez Gene: 8355 Human
    • Entrez Gene: 8356 Human
    • Entrez Gene: 8357 Human
    • Entrez Gene: 8358 Human
    • Entrez Gene: 8968 Human
    • Entrez Gene: 319152 Mouse
    • Entrez Gene: 319153 Mouse
    • Entrez Gene: 360198 Mouse
    • Entrez Gene: 97908 Mouse
    • Omim: 602810 Human
    • SwissProt: P68432 Cow
    • SwissProt: P68431 Human
    • SwissProt: P68433 Mouse
    • Unigene: 132854 Human
    • Unigene: 247813 Human
    • Unigene: 247814 Human
    • Unigene: 248176 Human
    • Unigene: 443021 Human
    • Unigene: 484990 Human
    • Unigene: 532144 Human
    • Unigene: 533292 Human
    • Unigene: 546315 Human
    • Unigene: 586261 Human
    • Unigene: 591778 Human
    • Unigene: 221301 Mouse
    • Unigene: 261657 Mouse
    • Unigene: 377874 Mouse
    • Unigene: 390558 Mouse
    • Unigene: 397328 Mouse
    see all
  • 别名

    • 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

图片

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)

    ab5819 staining Histone H3 (tri methyl K9, phospho S10) in mitotic HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab5819 at 0.5µg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). The secondary antibody (shown in green) was ab150081, Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h at room temperature. Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  • Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)
    Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)
    All lanes : Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819) at 1 µg/ml

    Lane 1 : Calf Thymus Histone at 0.5 µg
    Lane 2 : HeLa Nuclear – Triton Prep at 10 µg
    Lane 3 : NIH3T3 Nuclear – Triton Prep at 10 µg

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

    Predicted band size: 17 kDa
    Observed band size: 17 kDa


    Exposure time: 2 minutes


    Blocking buffer: 2% BSA

    Gel type: MES

  • Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)
    Western blot - Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819)
    All lanes : Anti-Histone H3 (tri methyl K9, phospho S10) antibody (ab5819) at 1/500 dilution

    Lane 1 : HeLa histones
    Lane 2 : Colcemid-treated HeLa histones
    Lane 3 : HeLa histones with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg
    Lane 4 : Colcemid-treated HeLa histones with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg
    Lane 5 : HeLa histones with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg
    Lane 6 : Colcemid-treated HeLa histones with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg
    Lane 7 : HeLa histones with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg
    Lane 8 : Colcemid-treated HeLa histones with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg
    Lane 9 : HeLa histones with Human Histone H3 (tri methyl K9) peptide (ab1773) at 1 µg
    Lane 10 : Colcemid-treated HeLa histones with Human Histone H3 (tri methyl K9) peptide (ab1773) at 1 µg

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 17 kDa
    Observed band size: 22 kDa why is the actual band size different from the predicted?
    Additional bands at: 60 kDa. We are unsure as to the identity of these extra bands.



    Rabbit polyclonal to Histone H3 (phospho S10, tri methyl K9) - ab5819, on histone preparations from HeLa cells. Primary antibody used at 1/500.

    Lanes 1, 3, 5, 7, 9 - control HeLa histones (0.5ug/lane)
    Lanes 2, 4, 6, 8, 10 - Colcemid-treated HeLa histones (0.5ug/lane)

    All blocking peptides are 1ug/lane

    Lanes 1 and 2: ab5819  
    Lanes 3 and 4: ab5819 + Histone H3 (Tri Me K9, phospho S10) peptide - ab15644
    Lanes 5 and 6: ab5819 + Histone H3 peptide (unmodified) - ab7228
    Lanes 7 and 8: ab5819 + Histone H3  (phospho S10) peptide - ab11477
    Lanes 9 and 10: ab5819 + Histone H3 (tri methyl K9) peptide - ab1773 

    Secondary antibody - Goat polyclonal to rabbit IgG (HRP) - ab6721 1/5000.

    ab5819 is specific for Histone H3 phosphorylated at residue S10 and tri methylated at residue K9. The activity of the antibody is blocked by the addition of the immunizing peptide, ab15644 (lanes 3 and 4).

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)This image is courtesy of Darin McDonald, Hendzel Laboratory

    SKN cells stained with ab5819 (green) at a dilution of 1/2000. The cells were fixed in paraformaldehyde for 10 minutes prior to incubation with ab5819. The DNA is stained with DAPI (blue). 100x magnification.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)

    ab5819 at a 1/500 dilution staining mitotic human HeLa cells by immunocytochemistry. The antibody was incubated with formaldehyde fixed cells for 1 1/2 hours and bound antibody was then detected using a Texas red conjugated goat anti-rabbit IgG.

    This image is courtesy of an Anonymous Abreview submitted on 6 April 2006.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade (ab5819)
    IHC image of Histone H3 (tri methyl K9, phospho S10) 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 ab5819, 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.

实验方案

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (14)

发表研究结果有使用 ab5819?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab5819 被引用在 14 文献中.

  • Calvet S  et al. Endogenous Retroviral Sequences Behave as Putative Enhancers Controlling Gene Expression through HP1-Regulated Long-Range Chromatin Interactions. Cells 11:N/A (2022). PubMed: 35954237
  • Yang L  et al. Bisphenol Exposure Disrupts Cytoskeletal Organization and Development of Pre-Implantation Embryos. Cells 11:N/A (2022). PubMed: 36291100
  • Cuddy SR  et al. Neuronal hyperexcitability is a DLK-dependent trigger of herpes simplex virus reactivation that can be induced by IL-1. Elife 9:N/A (2020). PubMed: 33350386
  • van Mierlo G  et al. Integrative Proteomic Profiling Reveals PRC2-Dependent Epigenetic Crosstalk Maintains Ground-State Pluripotency. Cell Stem Cell 24:123-137.e8 (2019). PubMed: 30472157
  • Suzuki T  et al. Arsenite-induced histone H3 modification and its effects on EGR1 and FOS expression in HeLa cells. J Appl Toxicol 38:734-743 (2018). PubMed: 29350772
View all Publications for this product

客户评价及客户问答

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提交评价 提交问题

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (tri methyl K9, phospho S10) antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Apr 06 2006

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

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