Human Histone H3 (tri methyl K27) peptide (ab1782)

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Dot Blot - Human Histone H3 (tri methyl K27) peptide (ab1782)

    Key features and details

    • Purity: > 70% 70 - 90% by HPLC
    • Suitable for: Dot blot, Blocking

    製品の詳細

    特性

    Our Abpromise guarantee covers the use of ab1782 in the following tested applications.

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

    • アプリケーション

      Dot blot

      Blocking - Blocking peptide for Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002), Anti-Histone H3 (di methyl K27, tri methyl K27) antibody [mAbcam 6147] (ab6147)

    • 製品の状態

      Liquid

    • 備考

      - First try to dissolve a small amount of peptide in either water or buffer. The more charged residues on a peptide, the more soluble it is in aqueous solutions.
      - If the peptide doesn’t dissolve try an organic solvent e.g. DMSO, then dilute using water or buffer.
      - Consider that any solvent used must be compatible with your assay. If a peptide does not dissolve and you need to recover it, lyophilise to remove the solvent.
      - Gentle warming and sonication can effectively aid peptide solubilisation. If the solution is cloudy or has gelled the peptide may be in suspension rather than solubilised.
      - Peptides containing cysteine are easily oxidised, so should be prepared in solution just prior to use.

    • Concentration information loading...

    前処理および保存

    • 保存方法および安定性

      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.

      Information available upon request.

    関連情報

    • 別名

      • H3 histone family member E pseudogene
      • H3 histone family, member A
      • H3/A
      • H31_HUMAN
      • H3F3
      • H3FA
      • Hist1h3a
      • HIST1H3B
      • HIST1H3C
      • HIST1H3D
      • HIST1H3E
      • HIST1H3F
      • HIST1H3G
      • HIST1H3H
      • HIST1H3I
      • HIST1H3J
      • HIST3H3
      • histone 1, H3a
      • Histone cluster 1, H3a
      • Histone H3 3 pseudogene
      • Histone H3.1
      • Histone H3/a
      • Histone H3/b
      • Histone H3/c
      • Histone H3/d
      • Histone H3/f
      • Histone H3/h
      • Histone H3/i
      • Histone H3/j
      • Histone H3/k
      • Histone H3/l
      see all

    • 機能

      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.

    • 配列類似性

      Belongs to the histone H3 family.

    • 発生段階

      Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.

    • 翻訳後修飾

      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.

    • 細胞内局在

      Nucleus. Chromosome.
    • Information by UniProt

    画像

    • Dot Blot - Human Histone H3 (tri methyl K27) peptide (ab1782)
      Dot Blot - Human Histone H3 (tri methyl K27) peptide (ab1782)This image is courtesy of an anonymous Abreview

      ab1782 (at 0.01 and 0.1 μg) used in dot blot to test for cross-reactivity of an anti-H3K27me3 polyclonal antibody (1/2500). A HRP-conjugated mouse anti-rabbit polyclonal (1/10000) was used as the secondary antibody.

      See Abreview

    プロトコール

    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

    参考文献 (9)

    ab1782 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab1782 は 9 報の論文で使用されています。

    • Kinkley S  et al. reChIP-seq reveals widespread bivalency of H3K4me3 and H3K27me3 in CD4(+) memory T cells. Nat Commun 7:12514 (2016). PubMed: 27530917
    • Maeng YS  et al. Histone methylation levels correlate with TGFBIp and extracellular matrix gene expression in normal and granular corneal dystrophy type 2 corneal fibroblasts. BMC Med Genomics 8:74 (2015). PubMed: 26553048
    • Hui SP  et al. Characterization of Proliferating Neural Progenitors after Spinal Cord Injury in Adult Zebrafish. PLoS One 10:e0143595 (2015). PubMed: 26630262
    • McLaughlin N  et al. In situ histone landscape of nephrogenesis. Epigenetics 9:222-35 (2014). PubMed: 24169366
    • Arambasic M  et al. Pdsg1 and Pdsg2, novel proteins involved in developmental genome remodelling in Paramecium. PLoS One 9:e112899 (2014). PubMed: 25397898
    • Huh YH & Sherley JL Decreased H3K27 and H3K4 trimethylation on mortal chromosomes in distributed stem cells. Cell Death Dis 5:e1554 (2014). PubMed: 25476902
    • Wang S  et al. Lineage specific trimethylation of H3 on lysine 4 during C. elegans early embryogenesis. Dev Biol 355:227-38 (2011). PubMed: 21549110
    • McManus KJ  et al. Dynamic changes in histone H3 lysine 9 methylations: Identification of a mitosis-specific function for dynamic methylation in chromosome congression and segregation. J Biol Chem : (2005). PubMed: 16373353
    • Chadwick BP & Willard HF Multiple spatially distinct types of facultative heterochromatin on the human inactive X chromosome. Proc Natl Acad Sci U S A : (2004). PubMed: 15574503

    レビューと Q&A

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    1-3 of 3 Abreviews or Q&A

    Question

    I am doing research with the model plant Arabidopsis. I can use the antibodies and peptides derived from human, because histone H3 is mostly conserved among many eukaryotes, however there are still differences at some amino acid positions. Therefore the information of the residue sequence or residue positions is very important for me to choose your products. In addition, the purity of the peptides is important to judge the quality of your products.

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    Answer

    The sequence and purification method is propriety, but if you let me know what is important for you I can check if one of our product fits.

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    Other (Dot Blot) Abreview for Histone H3 peptide - tri methyl K27

     Excellent
    Abreviews
    abreview image
    Application
    Other - Do not use

    Review text: I used this product to do a dot blot specificity test for cross reactivity of modified histone antibodies and everything worked great. I made two dilutions of the peptide and spotted it on a nitrocellulose membrane using a Bio-Dot apparatus. Once the peptides were bound to the membrane I incubated with the appropriate primary and secondary antibodies at an assay dependent concentration and developed using ECL.
    Sample: Human Purified protein

    Primary antibody (in addition to 'Histone H3 peptide - tri methyl K27')
    Name: Non-Abcam antibody was used: Anti-H3K27me3
    Host species: Rabbit
    Clonality: Polyclonal
    Dilution: 1/2500

    Secondary antibody
    Name: Non-Abcam antibody was used: Anti-rabbit
    Host species: Mouse
    Clonality: Polyclonal
    Conjugation: Horse Radish Peroxidase
    Dilution: 1/10000

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    Verified customer

    投稿 Oct 26 2010

    Question

    I am interested in doing some affinity purification of H3TrimK27 antibody to remove any antibodies that cross react with H3 trimethyl K9. Is there a way in which i can immobilize the H3TrimK27 peptide onto beads or are there sulfhybryl groups on the peptide?

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    Answer

    The sequence contains a cysteine residue on the end which we use to conjugate to the matrix. The matrix which we use is called Sulfolink coupling gel from Perbio (20402B). Alternatively NHS activated sepharose can be used which links via the primary amino group to the NHS (N-hydroxysuccinimide). This is supplied by Amersham. I hope this makes sense but I would recommend the sulfolink as it is a quick protocol. I think it is recommended to use a 1mg/ml peptide solution for optimal peptide binding.

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