Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915)

Overview

  • Product nameAnti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP GradeSee all Histone H3 primary antibodies ...
  • Description
    Rabbit polyclonal to Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) - ChIP Grade
  • SpecificityThis antibody recognizes histone H3 acetylated at lysines 9, 14, 18, 23 or 27 as confirmed by dot blot with nonmodified histone H3 peptide or peptides No reaction with non-modified histone H3 peptide as tested by dot blot.
  • Tested applicationsChIP, ICC/IF, WB more details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (Human) including acetyl-lysines contained in the N-terminal tail of human histone H3.

  • Positive control
    • HeLa cells. IF/ICC: HeLa cell line.

Properties

Applications

Our Abpromise guarantee covers the use of ab47915 in the following tested applications.

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

Application Notes
ChIP
ICC/IF
WB
  • Application notesChIP: Use at an assay dependent dilution (PMID 20080953).
    WB: 1/500 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • FunctionCore 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.
    • Sequence similaritiesBelongs to the histone H3 family.
    • Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
    • Post-translational
      modifications
      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.
    • Cellular localizationNucleus. Chromosome.
    • Target information above from: UniProt accession P68431 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links
    • Alternative names
      • H 3 antibody
      • H3 3 Like Sequence MH921 antibody
      • H3 3A antibody
      • H3 antibody
      • H3 Histone antibody
      • H3 Histone Family Member E Pseudogene antibody
      • H3.4 antibody
      • H3/A antibody
      • H3/g antibody
      • H31_HUMAN antibody
      • H3F3 antibody
      • H3FA antibody
      • H3FT antibody
      • H3t antibody
      • HIST1H3J antibody
      • HIST3H3 antibody
      • Histone cluster 1, H3a antibody
      • Histone H3 3 Pseudogene antibody
      • Histone H3.1 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
      see all

    Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade images

    • ICC/IF image of ab47915 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47915,1/1000 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG(H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Histone H3ac (pan-acetyl) antibody tested by ChIP. ChIP was performed using a High Sensitivity Kit with 5 µg of chromatin from HeLa cells and 10 µl of Histone H3ac antibody. ChIP DNA was used in qPCR with the negative control primer pairs or gene-specific primer pairs as indicated. Data are presented as Binding Events Detected per 1000 Cells using a normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

    • All lanes : Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915) at 1/2000 dilution

      Lane 1 : HeLa cells, no treatment
      Lane 2 : HeLa cells, treatment with sodium butyrate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 15 kDa
      Observed band size : 17 kDa (why is the actual band size different from the predicted?)

    References for Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915)

    This product has been referenced in:
    • Carvalho S  et al. Histone methyltransferase SETD2 coordinates FACT recruitment with nucleosome dynamics during transcription. Nucleic Acids Res 41:2881-93 (2013). WB ; Human . Read more (PubMed: 23325844) »
    • Yang LH  et al. Axin gene methylation status correlates with radiosensitivity of lung cancer cells. BMC Cancer 13:368 (2013). Read more (PubMed: 23915259) »

    See all 5 Publications for this product

    Product Wall



    I asked the lab what they used for the testing of this antibody and recieved the following answer:

    "The ChIP was made with the equivalent of 1.5 x 106 cells, this is approximately 10 to 15 ug of chromatin per 10ul antibody.&quo...

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    I am very pleased to hear you would like to accept our offer and test ab47915 in mouse. This code will give you $349 off your next order before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the ...

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    I can confirm that ab47915 antibody is sold as whole antiserum. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datashe...

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