Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)

Reviews (18)Q&A (2)References (99)
ChIP - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
  • Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
  • Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
  • Immunoprecipitation - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)

Key features and details

  • Rabbit polyclonal to Histone H3 (mono methyl K9) - ChIP Grade
  • Suitable for: IP, WB, ChIP, ICC/IF
  • Reacts with: Cow, Human, Arabidopsis thaliana
  • Isotype: IgG

Overview

  • Product name

    Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade
    See all Histone H3 primary antibodies

  • Description

    Rabbit polyclonal to Histone H3 (mono methyl K9) - ChIP Grade

  • Host species

    Rabbit

  • Specificity

    Weak cross reactivity is observed with mono methyl K27 Histone H3. No cross-reactivity is seen with di or tri methyl K27.

  • Tested applications

    Suitable for: IP, WB, ChIP, ICC/IFmore details

  • Species reactivity

    Reacts with: Cow, Human, Arabidopsis thaliana
    Predicted to work with: Mouse, Rat, Saccharomyces cerevisiae, Xenopus laevis, Indian muntjac, Schizosaccharomyces pombe, Mammals

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab1771)

  • General notes

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab9045 in the following tested applications.

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

Application Abreviews Notes
IP
Use at an assay dependent concentration.
WB (7)
1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 17 kDa). Can be blocked with Histone H3 peptide - mono methyl K9 (ab1771).
ChIP (1)
Use 4-5µg for 106 cells.
ICC/IF (4)
Use at an assay dependent concentration.
Notes
IP
Use at an assay dependent concentration.
WB
1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 17 kDa). Can be blocked with Histone H3 peptide - mono methyl K9 (ab1771).
ChIP
Use 4-5µg for 106 cells.
ICC/IF
Use at an assay dependent concentration.

Target

  • Function

    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.

  • Sequence similarities

    Belongs to the histone H3 family.

  • Developmental stage

    Expressed 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 localization

    Nucleus. Chromosome.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 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

Images

  • ChIP - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
    ChIP - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)

    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 ab9045 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)This image is courtesy of Kirk McManus in the lab of Michael Hendzel, Univeristy of Alberta

    Anti-mono methyl lysine 9 of histone H3 (green) has a distribution often associated with euchromatic probes (small foci).  Most of these foci localize to regions that contain obvious enrichments of DNA with DAPI staining (red).  The perinucleolar chromatin is typically a site enriched in monomethylated lysine 9. 

    Top left: Mono-methyl Lys 9 (ab9045); Bottom left: DAPI; Top right: Merge of ab9045 (green) and DAPI (red).

  • Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
    Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
    All lanes : Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045) at 1 µg/ml

    Lane 2 : Human Histone H3 (unmodified) peptide (ab7228)
    Lane 3 : Human Histone H3 (mono methyl K27) peptide (ab1780)
    Lane 4 : Human Histone H3 (di methyl K27) peptide (ab1781)
    Lane 5 : Human Histone H3 (tri methyl K27) peptide (ab1782)
    Lane 6 : Human Histone H3 (mono methyl K4) peptide (ab1340)
    Lane 7 : Human Histone H3 (mono methyl K9) peptide (ab1771)

    Predicted band size: 17 kDa



    Rabbit polyclonal to Histone H3 K9 Methyl K9 (1/1000)

    Peptides at 1 ug/ml

    1XTBS, 5%BSA, 0.5% Tween

    This antibody shows significantly greater reactivity with mono methyl K9. This can be seen in lane 7, as the addition of ab1771 (mono methyl K9) completely blocks the activity of ab9045. Weaker cross-reactivity is seen against mono methyl K27. This is shown in lane 3, as the addition of ab1780 only partially blocks the activity of ab9045.

  • Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
    Western blot - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
    All lanes : Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (unmodified) peptide (ab7228) at 0.5 µg/ml
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
    Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml
    Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg/ml
    Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg/ml
    Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with Human Histone H3 (tri methyl K27) peptide (ab1782) at 0.5 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 17 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)

    ab9045 staining Histone H3 (mono methyl K9) in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Triton for 1h. The cells were then incubated with the antibody ab9045 at 0.1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • Immunoprecipitation - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)
    Immunoprecipitation - Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)

    Histone H3 (mono methyl K9) was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Histone H3 (mono methyl K9) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab9045.
    Secondary: Anti-rabbit IgG VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution.
    Band: 17kDa: Histone H3 (mono methyl K9).

References (99)

Publishing research using ab9045? Please let us know so that we can cite the reference in this datasheet.

ab9045 has been referenced in 99 publications.

  • Sak A  et al. Chaetocin induced chromatin condensation: effect on DNA repair signaling and survival. Int J Radiat Biol 97:494-506 (2021). PubMed: 33428851
  • Bukova I  et al. Loss of Wiz Function Affects Methylation Pattern in Palate Development and Leads to Cleft Palate. Front Cell Dev Biol 9:620692 (2021). PubMed: 34150743
  • Neupert J  et al. An epigenetic gene silencing pathway selectively acting on transgenic DNA in the green alga Chlamydomonas. Nat Commun 11:6269 (2020). PubMed: 33293544
  • Tang DE  et al. Histone Demethylase JMJD1A Promotes Tumor Progression via Activating Snail in Prostate Cancer. Mol Cancer Res 18:698-708 (2020). PubMed: 32019811
  • Rowbotham K  et al. Differential SP1 interactions in SV40 chromatin from virions and minichromosomes. Virology 548:124-131 (2020). PubMed: 32838933
View all Publications for this product

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ChIP abreview for Anti-Histone H3 (methylated) antibody - ChIP Grade

 Excellent
Abreviews
abreview image
Application
ChIP
Sample
Rat Cell lysate - whole cell (Normal Liver and liver cancer cells)
Specification
Normal Liver and liver cancer cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step
Real-time PCR
Positive control
GAPDH
Negative control
Normal IgG
Read More

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Verified customer

Submitted Jul 10 2009

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