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Epigenetics and Nuclear Signaling Histones H3 Methylated
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RecombinantRabMAb

Recombinant Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)

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ChIP - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
  • Flow Cytometry - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
  • Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y49] to Histone H3 (di methyl K9) - BSA and Azide free
  • Suitable for: WB, Flow Cyt, ICC/IF, ChIP
  • Reacts with: Mouse, Rat, Human

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

View more associated products

Overview

  • Product name

    Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free
    See all Histone H3 primary antibodies
  • Description

    Rabbit monoclonal [Y49] to Histone H3 (di methyl K9) - BSA and Azide free
  • Host species

    Rabbit
  • Specificity

    The antibody only detects Histone H3 dimethylated on Lysine 9.

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, ChIPmore details
    Unsuitable for: IHC or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (di methyl K9). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa cell lysate. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. ChIP: Chromatin prepared from HeLa cells.
  • General notes

    Ab173325 is the carrier-free version of ab32521. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab173325 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y49
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Histones
    • H3
    • Methylated

Associated products

  • Alternative Versions

    • Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)
  • Compatible Secondaries

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

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

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

Applications

Our Abpromise guarantee covers the use of ab173325 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 Use at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC or IP.
  • 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: 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 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
      ChIP - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)

      Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab32521 (blue), and 20µl 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32521).
    • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
      Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)

      Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling Histone H3 (di methyl K9) with purified ab32521 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

      Control: PBS only

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32521).
    • Flow Cytometry - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
      Flow Cytometry - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)

      Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Histone H3 (di methyl K9) with purified ab32521 at 1/220 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32521).
    • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
      Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada

      ab32521 (1/500) staining Histone H3 di-methyl K9 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details, please refer to Abreview.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32521).
    • Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)
      Anti-Histone H3 (di methyl K9) antibody [Y49] - BSA and Azide free (ab173325)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (0)

    Publishing research using ab173325? Please let us know so that we can cite the reference in this datasheet.

    ab173325 has not yet been referenced specifically in any publications.

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