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Anti-Histone H3.3 (mutated G34W) 抗体 [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

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ChIP - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
  • Flow Cytometry - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
  • Dot Blot - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
  • Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23581-39] to Histone H3.3 (mutated G34W) - ChIP Grade – BSA and Azide free
  • Suitable for: ICC/IF, Flow Cyt, IHC-P, Dot blot, WB, ChIP
  • Reacts with: Human

製品の概要

  • 製品名

    Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free
    Histone H3.3 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR23581-39] to Histone H3.3 (mutated G34W) - ChIP Grade – BSA and Azide free

  • 由来種

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    See all applications and species data

  • 免疫原

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

  • ポジティブ･コントロール

    • WB: HEK-293T transfected with Histone H3.3 G34W expression vector containing a myc-His-tag, whole cell lysate. IHC-P: Human giant tumor of bone. Flow Cyt: 293T cells transfected with myc-tagged Histone H3.3 G34W construct. ICC/IF: 293T cells transfected with Histone H3.3 G34W-Myc plasmid.

  • 特記事項

    ab272700 is the carrier-free version of ab272691. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • バッファー

    pH: 7.2
    Constituent: PBS

  • キャリア・フリー

    はい
  • Concentration information loading...

  • 精製度

    Protein A purified

  • ポリ/モノ

    モノクローナル

  • クローン名

    EPR23581-39

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab272700 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

アプリケーション Species
ChIP
Human
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
アプリケーション Abreviews 特記事項
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Dot blot
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 15 kDa.
ChIP
Use at an assay dependent concentration.
特記事項
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Dot blot
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 15 kDa.
ChIP
Use at an assay dependent concentration.

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

    画像

    • ChIP - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
      ChIP - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

      Chromatin was prepared from 293T transfected with myc-His tagged Histone H3.3 mutated G34W and Histone H3.3 WT cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with formaldehyde for 10min.
      The ChIP was performed with 25 µg of chromatin, 2 µg of ab272691 (red), or 2 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
      Primers and probes are located in the first kb of the transcribed region.

      *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272691).

    • Flow Cytometry - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
      Flow Cytometry - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

      Flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized 293T (Human embryonic kidney epithelial cell) transfected with myc tagged Histone H3.3 WT construct (Left panel) and myc-tagged Histone H3.3 G34W construct (Right panel) cells labelling Histone H3.3(mutated G34 W) with ab272691 at 1/50 dilution (1µg). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272691).

    • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
      Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T cells labelling Histone H3.3(mutated G34 W) with ab272691 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing nuclear staining in 293T cells transfected with Histone H3.3 G34W-Myc plasmid, while no staining in 293T cells transfected with H3.3 WT -Myc plasmid. Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 µg/ml dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272691).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

      Immunohistochemical analysis of paraffin-embedded human giant cell tumor of bone tissue labeling Histone H3.3(mutated G34 W) with ab272691 at 1/250 dilution followed by ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human giant cell tumor of bone (PMID: 29757500). Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272691).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

      Immunohistochemical analysis of paraffin-embedded human chondroblastoma tissue labeling Histone H3.3(mutated G34 W) with ab272691 at 1/250 dilution followed by ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Negative control: No staining in human chondroblastoma (PMID: 29757500).

      Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272691).

    • Dot Blot - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
      Dot Blot - Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

      Dot blot analysis of Histone H3.3 (mutated G34 W) labeled with ab272691 at 1/1000 dilution.

      Lane 1: Histone H3.3 H3G34W peptide (aa28-37).
      Lane 2: Histone H3.3 H3G34W peptide (aa33-43).
      Lane 3: Histone H3.3 H3G34W peptide (aa28-43).
      Lane 4: Histone H3.3 WT peptide (aa28-43).

      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab272691).

    • Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)
      Anti-Histone H3.3 (mutated G34W) antibody [EPR23581-39] - ChIP Grade – BSA and Azide free (ab272700)

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (0)

    ab272700 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab272700 は論文での使用が確認できていません。

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