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  1. Link

    alexa-fluor-647-histone-h3-di-methyl-k9-antibody-mabcam-1220-ab203851.pdf

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Epigenetics and Nuclear Signaling Histones H3 Methylated
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Alexa Fluor® 647 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab203851)

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab203851)

    Key features and details

    • Alexa Fluor® 647 Mouse monoclonal [mAbcam 1220] to Histone H3 (di methyl K9)
    • Suitable for: ICC/IF
    • Reacts with: Human
    • Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm
    • Isotype: IgG2a

    Conjugates logo Related conjugates and formulations

    Alexa Fluor® 488 HRP Unconjugated

    You may also be interested in

    Peptide
    Product image
    Human Histone H3 (tri methyl K27) peptide (ab1782)
    Protein
    Product image
    Recombinant human Histone H3 protein (Active) (ab198757)
    Primary
    Product image
    Alexa Fluor® 488 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab203850)

    View more associated products

    Overview

    • Product name

      Alexa Fluor® 647 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220]
      See all Histone H3 primary antibodies
    • Description

      Alexa Fluor® 647 Mouse monoclonal [mAbcam 1220] to Histone H3 (di methyl K9)
    • Host species

      Mouse
    • Conjugation

      Alexa Fluor® 647. Ex: 652nm, Em: 668nm
    • Tested applications

      Suitable for: ICC/IFmore details
    • Species reactivity

      Reacts with: Human
    • Immunogen

      Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
      (Peptide available as ab1772)

    • Positive control

      • ICC/IF: HeLa cells.
    • General notes

      Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

      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. Avoid freeze / thaw cycle. Store In the Dark.
    • Storage buffer

      pH: 7.40
      Preservative: 0.02% Sodium azide
      Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
    • Concentration information loading...
    • Purity

      IgG fraction
    • Clonality

      Monoclonal
    • Clone number

      mAbcam 1220
    • Isotype

      IgG2a
    • Light chain type

      kappa
    • Research areas

      • Epigenetics and Nuclear Signaling
      • Histones
      • H3
      • Methylated

    Associated products

    • Alternative Versions

      • Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
      • Alexa Fluor® 488 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab203850)
      • HRP Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab62168)
    • Immunizing Peptide (Blocking)

      • Human Histone H3 (di methyl K9) peptide (ab1772)
    • Recombinant Protein

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

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab203851 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
    ICC/IF
    1/100.

    This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min).

    Notes
    ICC/IF
    1/100.

    This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min).

    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
      • Omim: 602810 Human
      • SwissProt: P68431 Human
      • 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
      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

    • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab203851)
      Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (ab203851)

      ab203851 staining Histone H3 (di methyl K9) in HeLa cells. The cells were fixed with 100% methanol (5 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-Tween for 1h. The cells were then incubated overnight at +4°C with ab203851 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

      This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).

    Protocols

    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (1)

    Publishing research using ab203851? Please let us know so that we can cite the reference in this datasheet.

    ab203851 has been referenced in 1 publication.

    • Dorosz J  et al. Structure-Based Design of a New Scaffold for Cell-Penetrating Peptidic Inhibitors of the Histone Demethylase PHF8. Chembiochem 18:1369-1375 (2017). PubMed: 28430394

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Flow Cytometry abreview for Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] (Alexa Fluor® 647)

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Drosophila melanogaster Cell (Drosophila larval brain)
    Permeabilization
    Yes - Triton X 100: 0.02%
    Gating Strategy
    all the cells
    Specification
    Drosophila larval brain
    Preparation
    Cell harvesting/tissue preparation method: larval brains were dissociated with collagenase 1hour at 30°C in Schneider medium 10%SVF
    Sample buffer: Schneider's Drosophila Medium
    Fixation
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 20 2018

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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