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Epigenetics and Nuclear Signaling Histones H3 Phosphorylated

Anti-Histone H3 (phospho S28)抗体(ab5169)

  • Datasheet
Reviews (3)Q&A (1)References (14)

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Western blot - Anti-Histone H3 (phospho S28) antibody (ab5169)
  • Peptide Array - Anti-Histone H3 (phospho S28) antibody (ab5169)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S28) antibody (ab5169)
  • Western blot - Anti-Histone H3 (phospho S28) antibody (ab5169)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S28) antibody (ab5169)

Key features and details

  • Rabbit polyclonal to Histone H3 (phospho S28)
  • Suitable for: WB, PepArr, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-Histone H3 (phospho S28)抗体
    参阅全部 Histone H3 一抗
  • 描述

    兔多克隆抗体to Histone H3 (phospho S28)
  • 宿主

    Rabbit
  • 特异性

    This antibody is specific for Histone H3 phosphorylated at residue Ser 28 and does not recognise the unmodified residue or another phosphorylated residue (Ser 10) on the same histone.
  • 经测试应用

    适用于: WB, PepArr, ICC/IFmore details
  • 种属反应性

    与反应: Human
    预测可用于: Drosophila melanogaster
  • 免疫原

    Synthetic peptide corresponding to Human Histone H3 aa 1-100 (phospho S28) conjugated to keyhole limpet haemocyanin.
    Database link: P68431
    (Peptide available as ab5499)

  • 常规说明

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as 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
    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

    • Epigenetics and Nuclear Signaling
    • Histones
    • H3
    • Phosphorylated

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 (phospho S10) peptide (ab11477)
  • Corresponding Unmodified Peptide

    • Human Histone H3 (unmodified ) peptide (ab2623)
  • Isotype control

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

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

应用

Our Abpromise guarantee covers the use of ab5169 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
PepArr Use a concentration of 0.02 - 0.002 µg/ml.
ICC/IF 1/5000.

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
    • Omim: 602810 Human
    • SwissProt: P68431 Human
    • 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
    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

  • Western blot - Anti-Histone H3 (phospho S28) antibody (ab5169)
    Western blot - Anti-Histone H3 (phospho S28) antibody (ab5169)
    Anti-Histone H3 (phospho S28) antibody (ab5169) at 1 µg/ml + Hela Whole Cell Lysate - Colcemid Treated at 2.5 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    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: 30 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab5169 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Peptide Array - Anti-Histone H3 (phospho S28) antibody (ab5169)
    Peptide Array - Anti-Histone H3 (phospho S28) antibody (ab5169)

    All batches of ab5169 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - phospho S28 peptide (ab5499), indicating that this antibody specifically recognises the Histone H3 - phospho S28 modification.

    ab17163 - Histone H3 unmodified

    ab5499 - Histone H3 - phospho S28

    ab11477 - Histone H3 - phospho S10

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S28) antibody (ab5169)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S28) antibody (ab5169)The image was submitted as part of a review by Krik McManus, University of British Columbia

    Indian Muntjac (top panel) and HeLa cells (bottom panel) immunofluorescently labelled with ab5169 (green) at a working dilution of 1/5000. The DNA is counterstained with DAPI and is shown in blue in the top panel and red in the bottom panel. This antibody gives a characteristic staining pattern for Histone H3 (phospho S28) whereby the signal increases in intensity during late G2 and continues to increase until metaphase. Upon entry into anaphase the signal begins to decrease until reaching basal levels by early G1. 100x magnification.

  • Western blot - Anti-Histone H3 (phospho S28) antibody (ab5169)
    Western blot - Anti-Histone H3 (phospho S28) antibody (ab5169)Image courtesy of Richelle Sopko, Harvard University, U.S.A.
    All lanes : Anti-Histone H3 (phospho S28) antibody (ab5169) at 1/1000 dilution

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

    Secondary
    All lanes : Donkey anti-rabbit IgG, Horseradish Peroxidase-L at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa


    Exposure time: 10 minutes


    0-4hr old wee shRNA embryos (lane 2) should display elevated phH3Ser28 levels relative to 0-4hr old EGFP shRNA embryos (lane 1). Blocked with 10% BSA.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S28) antibody (ab5169)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S28) antibody (ab5169)This image was submitted as part of a review by Kirk McManus, University of British Columbia

    In situ peptide competition was perfomed on paraformaldehyde-fixed HeLa cells. Four 25µl aliquots were made, to which 7.5µg (1.5µl) of no peptide, H3 unmodified peptide (ab2623), H3 phospho S10 peptide (ab11477) or H3 phospho S28 peptide (ab5499) was added, mixed by vortexing and incubated for 1 hour at room temperature. Cells on glass coverslips were fixed with 4% paraformaldehyde (10min) and gently washed twice with PBS, then permeabilized with 0.5% Triton X-100 in PBS (10min) and gently washed three times with PBS. The cells were immunofluorescently labeled with either the peptide-competed antibody or the control antibody (i.e. no peptide) for 30min at room temperature, washed briefly with PBS containing 0.1% Triton X-100 (1 min) and twice with PBS. The cells were then incubated with an appropriate dilution of a secondary antibody at room temperature for 30min, rinsed as above and mounted using a 90% glycerol in PBS mount

实验方案

  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

    • Datasheet
  • 文献 (14)

    发表研究结果有使用 ab5169?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab5169 被引用在 14 文献中.

    • Paul AS  et al. Co-option of Plasmodium falciparum PP1 for egress from host erythrocytes. Nat Commun 11:3532 (2020). PubMed: 32669539
    • Ganter M  et al. Plasmodium falciparum CRK4 directs continuous rounds of DNA replication during schizogony. Nat Microbiol 2:17017 (2017). PubMed: 28211852
    • Shandilya J  et al. Regulation of AURORA B function by mitotic checkpoint protein MAD2. Cell Cycle 15:2196-2201 (2016). PubMed: 27341405
    • Bhattacharya S  et al. Brief Communication: Featured Article: Histone H2A mono-ubiquitination and cellular transformation are inversely related in N-nitrosodiethylamine-induced hepatocellular carcinoma. Exp Biol Med (Maywood) 241:1739-44 (2016). PubMed: 27190257
    • Zhao L  et al. Original Research: Influence of okadaic acid on hyperphosphorylation of tau and nicotinic acetylcholine receptors in primary neurons. Exp Biol Med (Maywood) 241:1825-33 (2016). PubMed: 27190248
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-Histone H3 (phospho S28) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Dr. Richelle Sopko

    Verified customer

    提交于 Dec 17 2013

    Western blot abreview for Anti-Histone H3 (phospho S28) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    4.5 µg
    Gel Running Conditions
    Reduced Denaturing (12)
    Sample
    Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (Fruit fly embryo)
    Specification
    Fruit fly embryo
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    提交于 Nov 08 2013

    Question

    Inquiry: Im interested in follow H3S28phosphorilation by ChIP in Saccharomyces cerevisiae. Do you have any antibody? Does the H3S28ph (5169) works for yeast? and for ChIP analysis?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 13 2013

    Answer



    ab5169's immunogen shares 85% sequence homology with the yeast protein and another antibody, ab10543 shares between 76-92% homology with the yeast protein so both could work in yeast but this has not been tested and therefore we cannot guarantee this.

    Again neither of these antibodies have been tested in ChIP and so we cannot guarantee that these will work.

    If you wish to test one of these antibodies, I could give you a testing discount.

    Read More

    Abcam Scientific Support

    回复于 Feb 13 2013

    Immunocytochemistry abreview for Anti-Histone H3 (phospho S28) antibody

    Excellent
    Abreviews
    Abreviews
    Application
    Immunocytochemistry
    Sample
    Human Cell (HeLa and SK-N-SH)
    Specification
    HeLa and SK-N-SH
    Fixative
    Paraformaldehyde
    Read More

    Dr. Kirk Mcmanus

    Verified customer

    提交于 Oct 13 2005

    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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