RabMAb

Anti-Histone H3 (acetyl K27) 抗体 [EP16602] - ChIP Grade (ab177178)

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製品の概要

  • 製品名
    Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade
    Histone H3 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP16602] to Histone H3 (acetyl K27) - ChIP Grade
  • 由来種
    Rabbit
  • 特異性
    ab177178 binds K27ac alone and also when S28 is phosphorylated
  • アプリケーション
    適用あり: Flow Cyt, PepArr, IHC-P, WB, ICC/IF, ChIPmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human Histone H3 aa 1-100 (acetyl K27). The exact sequence is proprietary.
    Database link: P68431

  • ポジティブ･コントロール
    • WB: Untreated HeLa and treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates; untreated NIH/3T3 and treated with 7 mM Sodium butyrate for 24 hours whole cell lysates. IHC-P: Human colon, rat colon and mouse liver tissues. ICC/IF: HeLa cells. ChIP: Chromatin prepared from HeLa cells. Flow: HeLa cells
  • 特記事項

     

     

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

    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.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab177178 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
Flow Cyt 1/130.
PepArr Use at an assay dependent concentration.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/200. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
ICC/IF 1/8000.
ChIP Use 2 µg for 25 µg of chromatin.

ターゲット情報

  • 機能
    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.
  • 配列類似性
    Belongs to the histone H3 family.
  • 発生段階
    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • 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.
    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.
  • 細胞内局在
    Nucleus. Chromosome.
  • Information by UniProt
  • 参照データベース
    see all
  • 別名
    • 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

画像

  • Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    All lanes : Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178) at 1/20000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • ChIP - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    ChIP - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)

    Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab177178 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads as a control sample (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)

    Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H3 (acetyl K27) with ab177178 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human colon tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with Trichostatin A (500 ng/ml) for 4 hours or untreated, labeling Histone H3 (acetyl K27) with ab177178 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. Acetylation level increased after treatment with Trichostatin A (500 ng/ml) for 4 hours. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab177178 at 1/8000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution.

  • Flow Cytometry - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    Flow Cytometry - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)

    Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) treated (Red)/untreated (Green) with 500ng/ml Trichostatin A for 4 hours with purified ab177178 at 1/130 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

  • Peptide Array - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    Peptide Array - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)

    ab177178 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
     
    Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
     
    The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here.

  • Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    Western blot - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    All lanes : Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178) at 1/10000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysates
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 7 mM Sodium butyrate for 24 hours whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)

    Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H3 (acetyl K27) with ab177178 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat colon tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade (ab177178)

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Histone H3 (acetyl K27) with ab177178 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on mouse liver tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

参考文献

This product has been referenced in:
  • Mohaghegh N  et al. NextPBM: a platform to study cell-specific transcription factor binding and cooperativity. Nucleic Acids Res N/A:N/A (2019). Read more (PubMed: 30657937) »
  • Bandini C  et al. IRF4 Mediates the Oncogenic Effects of STAT3 in Anaplastic Large Cell Lymphomas. Cancers (Basel) 10:N/A (2018). Read more (PubMed: 29346274) »
See all 5 Publications for this product

ab177178 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。

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1-4 of 4 Abreviews or Q&A

Immunoprecipitation abreview for Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - nuclear (MDA-MB-231)
Total protein in input
1e+007 cells
Treatment
20ng/ml TNF alpha
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
MDA-MB-231
Read More

Ifeoluwa Adewumi

Verified customer

投稿 Feb 28 2017

Western blot abreview for Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Human Recombinant protein
Loading amount
0.25 µg
Blocking step
Milk as blocking agent for 11 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

投稿 Feb 28 2017

ChIP abreview for Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (MDA-MB-231)
Negative control
Normal rabbit IgG
Specification
MDA-MB-231
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Positive control
Input; 1A260 of nuclear lysate
Read More

Abcam user community

Verified customer

投稿 Feb 28 2017

Western blot abreview for Anti-Histone H3 (acetyl K27) antibody [EP16602] - ChIP Grade

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Human Cell lysate - nuclear (K-562)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
3 µg
Specification
K-562
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Dr. Ragnhild Eskeland

Verified customer

投稿 Jun 06 2016

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

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