Anti-Histone H3 (phospho S28)抗体[HTA28] (ab10543)

概述

  • 产品名称
    Anti-Histone H3 (phospho S28)抗体[HTA28]
    参阅全部 Histone H3 一抗
  • 描述
    大鼠单克隆抗体[HTA28] to Histone H3 (phospho S28)
  • 特异性
    Does not detect the unphosphorylated epitope. It detects the phosphorylated histone molecule at the onset of mitosis (prophase, metaphase and weaker at the beginning of anaphase), but not during late anaphase.
  • 经测试应用
    适用于: IHC-Fr, ICC/IF, WB, ICC, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Chicken, Hamster, Cow, Human, Drosophila melanogaster, Oikopleura, Common marmoset
  • 免疫原

    Synthetic peptide corresponding to Human Histone H3 aa 23-35 (phospho S28) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    KAARKSA PATGGV


    Database link: P68431

  • 阳性对照
    • Whole extract of cultured human acute T cell leukemia Jurkat cells, treated with Nocodazole.

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
    Preservative: 15mM Sodium Azide
    Constituents: 1% BSA, 0.01M PBS, pH 7.4
  • Concentration information loading...
  • 纯度
    Immunogen affinity purified
  • 纯化说明
    Purified from culture supernatant of hybridoma cells grown in a bioreactor
  • 克隆
    单克隆
  • 克隆编号
    HTA28
  • 同种型
    IgG2a
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab10543 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-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 2.5 µg/ml.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 15 kDa.

We advise you to centrifuge this product vial before use.

ICC Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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

图片

  • ab10543 staining Histone H3 (phospho S28) in Human neural progenitor cells from iPS cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 1% serum, 0.1% BSA in PBS for 30 minutes at room temperature. Samples were incubated with primary antibody (2ug/ml in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rat IgG2a polyclonal was used as the secondary antibody (1/500). Total cells were stained using DAPI (blue)

    See Abreview

  • All lanes : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 1 µg/ml

    Lane 1 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated
    Lane 2 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated with Human Histone H3 (unmodified ) peptide (ab2623) at 0.5 µg/ml
    Lane 3 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated with Human Histone H3 (phospho S28) peptide (ab5499) at 0.5 µg/ml
    Lane 4 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated with Human Histone H3 (phospho S10) peptide (ab11477) at 0.5 µg/ml

    Lysates/proteins at 2.5 µg per lane.

    Secondary
    Peroxidase Conjugated AffiniPure Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

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


    Exposure time : 20 minutes

    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 ab10543 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

  • ab10543 staining Histone H3 (phospho S28) in Mouse neural stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 4% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500 in PBS + 4% serum) for 16 hours at 4°C. An Alexa Fluor® 546-conjugated anti-rat polyclonal was used as the secondary antibody (1/500). DAPI is stainded blue

    See Abreview

  • ab10543 staining Histone H3 (phospho S28) in murine embryonic fibroblast cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in paraformaldehyde and permeabilised in -20°C ethanol. Samples were then incubated with primary antibody at a 1/200 dilution for 1 hour in 0.1%Saponin/ 2.5% BSA/ PBS. The secondary antibody used was a goat anti-rat IgG (H+L) conjugated to DyLight® 649 (red) used at a 1/300 dilution. Gamma tubulin (blue), Ki67 (green) using ab15580.
  • ab10543 staining HeLa cells by ICC/IF. The cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X100 in PBS. The cells were then stained with ab10543 at 1/1000 in PBS for 1h at 22°C. A goat anti-rat Alexa Fluro 488 (ab150157) at 1/200 was used as the secondary antibody. Nuclei are stained in red with DAPI. The antibody produces the expected mitotic-associated staining pattern and is extremely strong. MeOH fixed samples were also evaluated and produced a similar, strong staining pattern in mitotic cells.

  • ab10543 staining Histone H3 (phospho S28) in mouse embryonic brain tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with paraformaldehyde and blocking with 10% serum for 1 hour at 250C was performed. The sample was incubated with primary antibody (1/300)  at 40C for 16 hours. An Alexa Fluor®488-conjugated Goat polyclonal to rat IgG was used as secondary antibody at 1/500 dilution.

    See Abreview

文献

This product has been referenced in:
  • Matsuda Y  et al. Nestin phosphorylation at threonines 315 and 1299 correlates with proliferation and metastasis of human pancreatic cancer. Cancer Sci 108:354-361 (2017). Human . Read more (PubMed: 28002641) »
  • Sidhaye J & Norden C Concerted action of neuroepithelial basal shrinkage and active epithelial migration ensures efficient optic cup morphogenesis. Elife 6:N/A (2017). Zebrafish . Read more (PubMed: 28372636) »

See all 24 Publications for this product

客户评价及客户问答

Application
Immunohistochemistry (Frozen sections)
Sample
Petaurus breviceps Tissue sections (Skin tissue)
Permeabilization
Yes - Triton
Specification
Skin tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Oct 12 2017

Application
IHC - Wholemount
Sample
Drosophila melanogaster Embryo (wholemount)
Specification
wholemount
Username

Abcam user community

Verified customer

提交于 Jan 10 2017

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (human)
Specification
human
Permeabilization
No
Fixative
Parafilm
Username

Ms. Nikita Wright

Verified customer

提交于 Sep 29 2014

Application
Immunocytochemistry
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Mar 27 2014

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 Mar 04 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
1% serum, 0.1% triton, 0.1% BSA in PBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Sample
Human Cell (human neural progenitor cells from ips cell line)
Specification
human neural progenitor cells from ips cell line
Permeabilization
Yes - see blocking buffer
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jan 27 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde
Username

Dr. Kirk Mcmanus

Verified customer

提交于 Oct 03 2013

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. If you wish to test one of this antibody, I could give you a testing discount using our AbT...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Neonatal Heart)
Specification
Neonatal Heart
Fixative
Paraformaldehyde
Permeabilization
Yes - Saponin 0,1%
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 May 11 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse embryonic fibroblast (MEF))
Specification
mouse embryonic fibroblast (MEF)
Fixative
Paraformaldehyde
Permeabilization
Yes - -20C ethanol
Username

Abcam user community

Verified customer

提交于 Aug 19 2009

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