Anti-Histone H3 (acetyl K9) 抗体 - ChIP Grade (ab4441)

製品の概要

  • 製品名
    Anti-Histone H3 (acetyl K9) antibody - ChIP Grade
    Histone H3 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Histone H3 (acetyl K9) - ChIP Grade
  • 特異性
    In Dot blot detects 50ng of mono-acetylated peptide corresponding to position Lys9 in the N-terminal sequence of Histone H3. Does not detect the mono-acetylated peptide corresponding to acetyl-lysine at position 14 or unacetylated Histone H3.
  • アプリケーション
    適用あり: Dot blot, WB, ChIP, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Tetrahymena, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Toxoplasma gondii
    交差が予測される動物種: Saccharomyces cerevisiae
  • 免疫原

    Synthetic peptide corresponding to Human Histone H3 aa 1-12 (acetyl K9) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    ARTKQTAR(Ac)KSTG-C

  • ポジティブ・コントロール
    • WB: TSA-treated and acid extracted HeLa and NIH/3T3 cells. ICC:polytene chromosomes of Drosophila melanogaster

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab4441 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.

Detects 50ng of mono-acetylated peptide corresponding to position Lys9 in the N-terminal sequence of Histone H3. Does not detect the mono-acetylated peptide corresponding to acetyl-lysine at position 14 or unacetylated Histone H3.

WB 1/10000. Detects a band of approximately 17 kDa.
ChIP Use 2µg for 106 cells.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

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
    • H3.3A antibody
    • HIST1 cluster, H3E antibody
    • H3 histone family, member A antibody
    • H3.1 antibody
    • H3/l antibody
    • H3F3 antibody
    • H3FF antibody
    • H3FJ antibody
    • H3FL antibody
    • Histone gene cluster 1, H3 histone family, member E antibody
    • histone H3.1t antibody
    • Histone H3/o antibody
    • FLJ92264 antibody
    • H 3 antibody
    • H3 antibody
    • H3 histone family, member B antibody
    • H3 histone family, member C antibody
    • H3 histone family, member D antibody
    • H3 histone family, member F antibody
    • H3 histone family, member H antibody
    • H3 histone family, member I antibody
    • H3 histone family, member J antibody
    • H3 histone family, member K antibody
    • H3 histone family, member L antibody
    • H3 histone family, member T antibody
    • H3 histone, family 3A antibody
    • H3/A antibody
    • H3/b antibody
    • H3/c antibody
    • H3/d antibody
    • h3/f antibody
    • H3/h antibody
    • H3/i antibody
    • H3/j antibody
    • H3/k antibody
    • H3/t antibody
    • H31_HUMAN antibody
    • H3F1K antibody
    • H3F3A antibody
    • H3FA antibody
    • H3FB antibody
    • H3FC antibody
    • H3FD antibody
    • H3FH antibody
    • H3FI antibody
    • H3FK antibody
    • HIST1 cluster, H3A antibody
    • HIST1 cluster, H3B antibody
    • HIST1 cluster, H3C antibody
    • HIST1 cluster, H3D antibody
    • HIST1 cluster, H3F antibody
    • HIST1 cluster, H3G antibody
    • HIST1 cluster, H3H antibody
    • HIST1 cluster, H3I antibody
    • HIST1 cluster, H3J 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 1, H3b antibody
    • Histone 1, H3c antibody
    • Histone 1, H3d antibody
    • Histone 1, H3e antibody
    • Histone 1, H3f antibody
    • Histone 1, H3g antibody
    • Histone 1, H3h antibody
    • Histone 1, H3i antibody
    • Histone 3, H3 antibody
    • histone cluster 1 H3 family member a antibody
    • histone cluster 1 H3 family member b antibody
    • histone cluster 1 H3 family member c antibody
    • histone cluster 1 H3 family member d antibody
    • histone cluster 1 H3 family member e antibody
    • histone cluster 1 H3 family member f antibody
    • histone cluster 1 H3 family member g antibody
    • histone cluster 1 H3 family member h antibody
    • histone cluster 1 H3 family member i antibody
    • histone cluster 1 H3 family member j antibody
    • Histone cluster 1, H3a antibody
    • Histone cluster 1, H3b antibody
    • Histone cluster 1, H3c antibody
    • Histone cluster 1, H3d antibody
    • Histone cluster 1, H3e antibody
    • Histone cluster 1, H3f antibody
    • Histone cluster 1, H3g antibody
    • Histone cluster 1, H3i antibody
    • Histone cluster 1, H3j antibody
    • Histone gene cluster 1, H3 histone family, member A antibody
    • Histone gene cluster 1, H3 histone family, member B antibody
    • Histone gene cluster 1, H3 histone family, member C antibody
    • Histone gene cluster 1, H3 histone family, member D antibody
    • Histone gene cluster 1, H3 histone family, member F antibody
    • Histone gene cluster 1, H3 histone family, member G antibody
    • Histone gene cluster 1, H3 histone family, member H antibody
    • Histone gene cluster 1, H3 histone family, member I antibody
    • Histone gene cluster 1, H3 histone family, member J antibody
    • Histone gene cluster 1, H3A antibody
    • Histone gene cluster 1, H3B antibody
    • Histone gene cluster 1, H3C antibody
    • Histone gene cluster 1, H3D antibody
    • Histone gene cluster 1, H3E antibody
    • Histone gene cluster 1, H3F antibody
    • Histone gene cluster 1, H3G antibody
    • Histone gene cluster 1, H3H antibody
    • Histone gene cluster 1, H3I antibody
    • Histone gene cluster 1, H3J antibody
    • Histone H 3 antibody
    • Histone H3.1 antibody
    • Histone H3.2 antibody
    • Histone H3.3 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
    • Histone H3/m antibody
    see all

画像

  • Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab4441 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

  • ChIP analysis using unpurified ab4441 binding Histon H3 in human primary CD34+ cell lysate. Cells were cross-linked for 15 minutes with 1% formaldehyde. Samples were incubated with primary antibody (0.2 µg/µg of chromatin) for 16 hours at 4°C. Protein binding was detected using real-time PCR.

    Positive controls:
    Region upstream of the transcription start site of the ACTB gene.
    Region upstream of the transcription start site of the EGR1 gene.

    Negative Controls:
    Region on chromosome 12 (Untr12) that is far from any known gene annotation and not expected to be bound by Histone H3 (acetyl K9).

    See Abreview

  • All lanes : Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441) at 1/2500 dilution

    Lane 1 : Mouse MEL cell nuclear lysate
    Lane 2 : Mouse MEL cell nuclear lysate treated with 0.4 µM Trichostatin A treatment for 18 hours.

    Lysates/proteins at 9 µg per lane.

    Secondary
    Donkey Anti-Rabbit IgG H&L (HRP) (ab6802) at 1/20000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview.

    See Abreview

  • Fruit fly (Drosophila melanogaster) Cell (polytene chromosomes) were fixed in formaldehyde, blocked in 1% BSA for 30 minutes and incubated with ab4441 (1/100) for 12 hours.

     

  • All lanes : Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441) at 0.5 µg/ml

    Lane 1 : Untreated HeLa cell acid-extract
    Lane 2 : HeLa cell acid-extract treated with sodium butyrate

    ab4441, Histone H3, acetylated (Lys9) Pab Rabbit x Human

参考文献

This product has been referenced in:
  • Chakraborty K  et al. The mito-DAMP cardiolipin blocks IL-10 production causing persistent inflammation during bacterial pneumonia. Nat Commun 8:13944 (2017). Read more (PubMed: 28074841) »
  • Meisenberg C  et al. Epigenetic changes in histone acetylation underpin resistance to the topoisomerase I inhibitor irinotecan. Nucleic Acids Res 45:1159-1176 (2017). WB ; Human . Read more (PubMed: 28180300) »

See all 184 Publications for this product

レビューと Q&A

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - nuclear (U2OS)
Specification
U2OS
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 May 15 2014

Application
Western blot
Loading amount
1 µg
Gel Running Conditions
Reduced Denaturing (18 % SDS-PAGE)
Sample
Dictyostelium discoideum Purified protein (Vegetative cells)
Specification
Vegetative cells
Blocking step
No blocking step used for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Dr. Duen-Wei Hsu

Verified customer

投稿 Mar 31 2014

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Human Cell lysate - nuclear (Human HeLa cell)
Specification
Human HeLa cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Nov 05 2013

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
Specification
Mouse embryonic fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Oct 24 2013

Application
Western blot
Loading amount
3 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
Username

Abcam user community

Verified customer

投稿 Oct 21 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step
Enzymatic
Sample
Rat Tissue sections (GONAD)
Specification
GONAD
Permeabilization
No
Fixative
Formaldehyde
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投稿 Sep 20 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (DLD1 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
DLD1 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Feb 13 2017

Application
Western blot
Sample
Ferret Tissue lysate - whole (Brain tissue)
Gel Running Conditions
Reduced Denaturing (10% Bis-Tris gel, Invitrogen)
Loading amount
100 µg
Treatment
100-200 nM of Trichostatin A (TSA), HDAC inhibitor for 24 hours
Specification
Brain tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Francis Djankpa

Verified customer

投稿 Feb 01 2017

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Astatotilapia burtoni Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 10 min in 0.2% NP-40 in TBS
Specification
Brain
Blocking step
10% serum and 1% BSA in TBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Sebastian Alvarado

Verified customer

投稿 Dec 01 2015

Application
ChIP
Sample
Human Cell lysate - whole cell (HeLa cervical cancer)
Negative control
pseudogene/heterochromatin region of the genome
Specification
HeLa cervical cancer
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: 1% formaldehyde
Positive control
Actively transcribing region of genome
Username

Abcam user community

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投稿 Jun 15 2015

1-10 of 42 Abreviews or Q&A

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