Anti-Histone H3.3 (phospho S31) 抗体 [EPR1873] - ChIP Grade (ab92628)

製品の概要

  • 製品名Anti-Histone H3.3 (phospho S31) antibody [EPR1873] - ChIP Grade
    Histone H3.3 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR1873] to Histone H3.3 (phospho S31) - ChIP Grade
  • 特異性Detects Histone H3.3 when phosphorylated at S31.
  • アプリケーション適用あり: Dot blot, ChIP, WB, IHC-P, ICC/IFmore details
    適用なし: Flow Cyt or IP
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Histone H3.3 aa 1-100.
    Database link: P84243

  • ポジティブ・コントロール
    • Untreated Hela cells and HeLa cell lysate treated with Calyculin A and/or nocodazole. Human cervical carcinoma
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated "PUR" on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab92628 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Dot blot 1/1000.
ChIP Use at an assay dependent concentration. PubMed: 22649058
WB 1/1000 - 1/5000. Predicted molecular weight: 15 kDa.
IHC-P 1/100 - 1/250.
ICC/IF 1/100 - 1/250.
  • 追加情報Is unsuitable for Flow Cyt or IP.
  • ターゲット情報

    • 機能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.
    • 配列類似性Belongs to the histone H3 family.
    • 発生段階Expressed throughout the cell cycle independently of DNA synthesis.
    • 翻訳後修飾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.
    • 細胞内局在Nucleus. Chromosome.
    • Information by UniProt
    • 参照データベース
    • 別名
      • H3 histone family 3A antibody
      • H3 histone family 3B antibody
      • H3 histone, family 3B (H3.3B) antibody
      • H3.3 antibody
      • H3.3A antibody
      • H3.3B antibody
      • H33_HUMAN antibody
      • H3F3 antibody
      • H3F3A antibody
      • H3f3b antibody
      • Histone H3.3 antibody
      • Histone H3.3Q antibody
      • Histone H3.A antibody
      • Histone H3.B antibody
      • MGC87782 antibody
      • MGC87783 antibody
      see all

    Anti-Histone H3.3 (phospho S31) antibody [EPR1873] - ChIP Grade 画像

    • Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% tritonX-100 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, LP starved and non-starved, labeling anti-Histone H3.3 (phospho S31) with Ab92628 at 1/500 dilution followed by Goat anti-Rabbit secondary IgG AlexaFluor®488 (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing nuclear staining on M phase of HeLa cells, then the signal decreased after LP treatment.

      For the pan antibody, there was no great difference after LP treatment. The data showed mostly nuclear staining.

    • All lanes : Anti-Histone H3.3 (phospho S31) antibody [EPR1873] - ChIP Grade (ab92628) at 1/1000 dilution

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : Whole cell lysate from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 100ng/ml of nocodazole for 18 hours
      Lane 3 : Whole cell lysate from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 100ng/ml of nocodazole for 18 hours. Membrane incubated with phosphatase

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 15 kDa


      Exposure time : 15 seconds

      Blocking/dilution buffer: 2% BSA/TBST

    • Dot Blot analysis of Lane 1: Histone H3.3 (pS31) phospho peptide and Lane 2: Histone H3.3 non-phospho peptide labeling Histone H3.3 (phospho S31) with ab92628 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. 

      ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 10 seconds.

       

       

       

       

       

    • All lanes : Anti-Histone H3.3 (phospho S31) antibody [EPR1873] - ChIP Grade (ab92628) at 1/1000 dilution

      Lane 1 : HeLa cell lysate untreated
      Lane 2 : HeLa cell lysate treated with Calyculin A

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 15 kDa
    • Paraffin-embedded human cervical carcinoma labelled with ab92628 at 1/100 dilution.


    • Predicted band size : 15 kDa

      This image is courtesy of an anonymous Abreview

      Representative western blot detecting Histone H3.3 (phospho S31) using ab92628 at 1/3000 dilution.

      Recombinant Histone H3.3 protein was treated with recombinant kinase and 200 µM ATP for the indicated times. 0.2 µg of protein was loaded to each lane and phosphorylated Histone H3.3 was detected using ab92628. An HRP-conjugated goat anti-rabbit polyclonal (1/20000) was used as the secondary antibody

      See Abreview

    Anti-Histone H3.3 (phospho S31) antibody [EPR1873] - ChIP Grade (ab92628) 使用論文

    This product has been referenced in:
    • Kron P  et al. Hypoxia-driven Hif2a coordinates mouse liver regeneration by coupling parenchymal growth to vascular expansion. Hepatology 64:2198-2209 (2016). Read more (PubMed: 27628483) »
    • Yuen BT  et al. Histone H3.3 regulates dynamic chromatin states during spermatogenesis. Development 141:3483-94 (2014). Read more (PubMed: 25142466) »

    See all 4 Publications for this product

    Product Wall

    Application Western blot
    Sample Chicken Recombinant protein (DT40)
    Gel Running Conditions Reduced Denaturing (4-12)
    Loading amount 500000 cells
    Specification DT40
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Ms. Marina Romanello

    Verified customer

    投稿 Jun 02 2015

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
    Sample Mouse Cell (Testis)
    Specification Testis
    Permeabilization Yes - Triton X100
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 Dec 23 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Cell lysate - whole cell (Raw 264.7)
    Loading amount 20 µg
    Specification Raw 264.7
    Treatment DRB and Vehicle control 0 - 2hrs
    Gel Running Conditions Reduced Denaturing (10)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 23°C
    Username

    Abcam user community

    Verified customer

    投稿 Jul 31 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Recombinant protein (H3.3 recombinant protein)
    Loading amount 0.2 µg
    Specification H3.3 recombinant protein
    Treatment Kinase and 200uM ATP
    Gel Running Conditions Reduced Denaturing (10%)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    投稿 Jun 08 2012

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"