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Epigenetics and Nuclear Signaling Histones H3 Methylated
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Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)

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  • SDS
Reviews (85)Q&A (21)References (1218)

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ChIP - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
  • ChIP - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
  • Western blot - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
  • Immunocytochemistry - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)

Key features and details

  • Rabbit polyclonal to Histone H3 (tri methyl K9) - ChIP Grade
  • Suitable for: WB, IHC-P, ICC/IF, ChIP
  • Reacts with: Mouse, Cow, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade
    See all Histone H3 primary antibodies
  • Description

    Rabbit polyclonal to Histone H3 (tri methyl K9) - ChIP Grade
  • Host species

    Rabbit
  • Specificity

    Histone H3 (tri methyl K9) antibody (ab8898) is specific for Histone H3 tri methyl Lysine 9. Shows slight cross-reactivity with tri methyl K27, which shares a similar epitope (please see Western blot image). Does not react with mono or di methylated K9.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, ChIPmore details
  • Species reactivity

    Reacts with: Mouse, Cow, Human
    Predicted to work with: Rat, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Drosophila melanogaster, Indian muntjac, Mammals, Xenopus tropicalis, Cyanidioschyzon merolae
  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (N terminal) (tri methyl K9) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab1773)

  • Positive control

    • ChIP: U2OS cells, mouse ES cells. WB: Calf Thymus Histone Preparation Nuclear Lysate. IHC-P: Normal human colon. ICC: Mouse 3T3MEF, Indian muntjac fibroblast cells, HeLa cells, Mouse Embryonic Stem cells.
  • General notes

    Every new batch of ab8898 is tested in house in ChIP. Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.

    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

    • Epigenetics and Nuclear Signaling
    • Histones
    • H3
    • Methylated
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • ChIP Related Products

    • Anti-Histone H3 (tri methyl K9) antibody [EPR16601] - ChIP Grade (ab176916)
    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Control Peptide

    • Human Histone H3 (mono methyl K4) peptide (ab1340)
    • Human Histone H3 (tri methyl K4) peptide (ab1342)
    • Human Histone H3 (mono methyl K9) peptide (ab1771)
    • Human Histone H3 (di methyl K9) peptide (ab1772)
    • Human Histone H3 (mono methyl K27) peptide (ab1780)
    • Human Histone H3 (di methyl K27) peptide (ab1781)
    • Human Histone H3 (tri methyl K27) peptide (ab1782)
    • Human Histone H3 (unmodified) peptide (ab7228)
    • Human Histone H3 (di methyl K4) peptide (ab7768)
  • Corresponding Unmodified Peptide

    • Human Histone H3 (unmodified ) peptide (ab2903)
  • Immunizing Peptide (Blocking)

    • Human Histone H3 (tri methyl K9) peptide (ab1773)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • SAT-alpha positive control ChIP primer pair (ab269263)
  • Recombinant Protein

    • Recombinant human Histone H3 protein (Active) (ab198757)
  • Related Products

    • Histone H3 (K9) Methyltransferase Activity Quantification Assay Kit (ab113453)
    • Histone H3 (tri-methyl K9) Quantification Kit (Colorimetric) (ab115064)
    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
    • Anti-Histone H3 (tri methyl K9) antibody [6F12-H4] (ab184677)
    • Anti-Histone H3 (tri methyl K9) antibody [EPR16601] - BSA and Azide free (ab232324)
    • Human Histone H3 (di methyl K4) peptide (ab7768)
    • Anti-Histone H3 (mono methyl K9) antibody - ChIP Grade (ab9045)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab8898 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
WB (29)
Use at an assay dependent concentration. Predicted molecular weight: 15 kDa.Can be blocked with Human Histone H3 (tri methyl K9) peptide (ab1773).
IHC-P (8)
1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF (18)
1/500.
ChIP (25)
Use 2-4 µg for 25 µg of chromatin.

We recommend SAT-alpha ChIP primer pair ab269263 as a positive control.

Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 15 kDa.Can be blocked with Human Histone H3 (tri methyl K9) peptide (ab1773).
IHC-P
1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
1/500.
ChIP
Use 2-4 µg for 25 µg of chromatin.

We recommend SAT-alpha ChIP primer pair ab269263 as a positive control.

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.
  • 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

    • Entrez Gene: 8350 Human
    • Entrez Gene: 8351 Human
    • Entrez Gene: 8352 Human
    • Entrez Gene: 8353 Human
    • Entrez Gene: 8354 Human
    • Entrez Gene: 8355 Human
    • Entrez Gene: 8356 Human
    • Entrez Gene: 8357 Human
    • Entrez Gene: 8358 Human
    • Entrez Gene: 8968 Human
    • Entrez Gene: 319152 Mouse
    • Entrez Gene: 319153 Mouse
    • Entrez Gene: 360198 Mouse
    • Entrez Gene: 97908 Mouse
    • Entrez Gene: 100364501 Rat
    • Entrez Gene: 100365669 Rat
    • Entrez Gene: 291159 Rat
    • Entrez Gene: 679994 Rat
    • Entrez Gene: 680511 Rat
    • Entrez Gene: 682330 Rat
    • Omim: 602810 Human
    • SwissProt: P68432 Cow
    • SwissProt: P68431 Human
    • SwissProt: P68433 Mouse
    • SwissProt: Q6LED0 Rat
    • Unigene: 132854 Human
    • Unigene: 247813 Human
    • Unigene: 247814 Human
    • Unigene: 248176 Human
    • Unigene: 443021 Human
    • Unigene: 484990 Human
    • Unigene: 532144 Human
    • Unigene: 533292 Human
    • Unigene: 546315 Human
    • Unigene: 586261 Human
    • Unigene: 591778 Human
    • Unigene: 221301 Mouse
    • Unigene: 261657 Mouse
    • Unigene: 377874 Mouse
    • Unigene: 390558 Mouse
    • Unigene: 397328 Mouse
    • Unigene: 138090 Rat
    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 (tri methyl K9) antibody - ChIP Grade (ab8898)
    ChIP - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)

    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  ab8898 (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.

  • ChIP - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    ChIP - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)This image is courtesy of an anonymous abreview.

    X-Chip assay was performed using nuclear lysates prepared from mouse ES cells. Crosslinking was done for 15 minutes in 1% formaldehyde. Primary antibody was incubated first with peptides ab7228, ab1342, ab1782, ab1773, ab1772 and ab1771 in a chip competition assay and then used in chip at 0.0133µg/ µg chromatin (chip sonication buffer) and incubated with sample for 24 hours at 4°C.

    Positive control: ChIP coupled with a peptide competition assay to validate the specificity of the antibody.

    Negative control: Genomic region (chr10:79154149-79155200) with no evidence of H3K9me3. 

    RT-PCR detection method was used.

    Polrmt: PCR primers situated in the coding regions of Polymerase (RNA) mitochondrial (DNA directed).

    Agrn: PCR primers situated in the coding regions of Agrin

  • Western blot - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    Western blot - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    All lanes : Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified) peptide (ab7228) at 0.5 µg/ml
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 0.5 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 0.5 µg/ml
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 0.5 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 0.5 µg/ml
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml
    Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K9) peptide (ab1773) at 0.5 µg/ml
    Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K27) peptide (ab1780) at 0.5 µg/ml
    Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K27) peptide (ab1781) at 0.5 µg/ml
    Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate 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: 15 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?



    Lane 8 shows that Rabbit polyclonal to Histone H3 (tri methyl K9) is blocked by the addition of the immunizing peptide (ab1773). Cross-reactivity with Histone H3 peptide - tri methyl K27 (ab1782) is also shown in Lane 11.
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)This image is a courtesy of Anonymous Abreview

    ab8898 staining H3K9me3 in mouse 3T3MEF by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with PFA, permeabilized with CSK buffer and MeOH and blocked with 5% BSA for 30 minutes at 21°C. Samples were incubated with primary antibody (1/500 in PBS +5% BSA + 01% Tween20) for 2 hours at 21°C. An undiluted Alexa Fluor® 546-conjugated Goat anti-rabbit IgG was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    Immunocytochemistry - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)

    ab8898 staining Histone H3 (tri methyl K9) in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab8898 at 0.5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Also suitable in cells fixed with 4% paraformaldehyde (10 min).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)

    IHC image of ab8898 staining Histone H3 (tri methyl K9) in normal human colon formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8898, 1/400 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    These images were kindly submitted by Prof Bryan Turner, University of Birmingham. Undifferentiated Mouse Embryonic Stem cells or cells differentiated for 7 days were incubated with ab8898. The staining is specific for centromeric heterochromatin on metaphase chromosomes.
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)This image is courtesy of Kirk McManus in the lab of Michael Hendzel, Univeristy of Alberta

    Indian muntjac fibroblast cells stained with anti-Histone H3 tri methyl K9, ab8898, (green, left panel, deconvolution image; red, right panel, epifluorescence image).

    The centromeres are enriched in Histone H3 tri methyl K9. There are also additional bands that occur throughout the chromosomes. Note that these images are taken in situ and are imaged under conditions where distinct cytogenetic-like banding patterns have not previously been possible to visualize (e.g., several acetylated antibodies have been reported to be associated with chromosome bands but, although not homogenously distributed along in situ chromosomes, they do not generate distinct banding patterns).

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade (ab8898)

    ICC/IF image of ab8898 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 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 (ab8898, 0.1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, a goat anti-rabbit DyLight® 488 (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.

Protocols

  • Chromatin Immunofluorescence

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (1218)

Publishing research using ab8898? Please let us know so that we can cite the reference in this datasheet.

ab8898 has been referenced in 1218 publications.

  • Zhang J  et al. Down-regulation of Suv39h1 attenuates neointima formation after carotid artery injury in diabetic rats. J Cell Mol Med 24:973-983 (2020). PubMed: 31736204
  • Yin N  et al. IDH1-R132H mutation radiosensitizes U87MG glioma cells via epigenetic downregulation of TIGAR. Oncol Lett 19:1322-1330 (2020). PubMed: 31966064
  • Ormundo LF  et al. LINE-1 specific nuclear organization in mice olfactory sensory neurons. Mol Cell Neurosci 105:103494 (2020). PubMed: 32387751
  • Gylling HM  et al. Repeat RNAs associate with replication forks and post-replicative DNA. RNA N/A:N/A (2020). PubMed: 32393525
  • González B  et al. Dopamine Receptor D1 Contributes to Cocaine Epigenetic Reprogramming of Histone Modifications in Male Germ Cells. Front Cell Dev Biol 8:216 (2020). PubMed: 32318569
View all Publications for this product

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HBE)
Permeabilization
Yes - 0.5% Triton in PBS
Specification
HBE
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

Submitted Mar 05 2021

Western blot abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
30 µg
Specification
Heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

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Submitted Aug 17 2020

ChIP abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Mouse Cell lysate - other (mouse ES cells, 300,000 cells, digitonin 0.03% in)
Specification
mouse ES cells, 300,000 cells, digitonin 0.03% in
Detection step
Other
Type
Native ChIP (N-ChIP)
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Abcam user community

Verified customer

Submitted Feb 03 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Tammar wallaby Cell (ovary)
Permeabilization
Yes - 0.1% (v/v) Triton X-100 in PBS
Specification
ovary
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 27°C
Fixative
Paraformaldehyde
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Verified customer

Submitted Mar 13 2019

Western blot abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - nuclear (MEF)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
5 µg
Specification
MEF
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Jan 23 2019

CHIPseq abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
ChIP-sequencing
Sample
Mouse Tissue lysate - whole (Brain)
Specification
Brain
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Verified customer

Submitted Jul 05 2018

ChIP abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Mouse Tissue lysate - whole (Brain)
Negative control
Arc gene (highly expressed in brain tissue), promoter region
Specification
Brain
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Positive control
Gcg gene (glucagon gene, highly repressed in brain tissue), promoter region Hbbb1 gene (hemoglobin B1 subunit gene, highly repressed in brain tissue), promoter region
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Verified customer

Submitted Jun 18 2018

Western blot abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (B Cell lymphoma)
Gel Running Conditions
Reduced Denaturing (4-20% Gradient)
Loading amount
30 µg
Specification
B Cell lymphoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Nov 13 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Arvicanthis niloticus (African grass rat) Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA buffer
Permeabilization
Yes - Apply PBS with 0.3% Triton X-100 for 15 min
Specification
Liver
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formalin
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Hui Huang

Verified customer

Submitted Oct 04 2017

ChIP abreview for Anti-Histone H3 (tri methyl K9) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - whole cell (Human Epithelial OVCAR cell line)
Negative control
IgG
Specification
Human Epithelial OVCAR cell line
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 (1%)
Positive control
-
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Abcam user community

Verified customer

Submitted Jul 05 2017

1-10 of 106 Abreviews or Q&A

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