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  1. Link

    human-histone-h3-di-methyl-k4-peptide-ab7768.pdf

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Epigenetics and Nuclear Signaling Histones H3 Methylated
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Human Histone H3 (di methyl K4) peptide (ab7768)

  • Datasheet
Reviews (1)Q&A (5)References (6)

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Dot Blot - Human Histone H3 (di methyl K4) peptide (ab7768)

    Key features and details

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

    You may also be interested in

    Conjugation
    Product image
    Streptavidin Conjugation Kit - Lightning-Link® (ab102921)
    Peptide
    Human Histone H4 (acetyl K12) peptide (ab15662)
    Primary
    Product image
    Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)

    View more associated products

    Description

    • Product name

      Human Histone H3 (di methyl K4) peptide
      See all Histone H3 proteins and peptides
    • Purity

      > 90 % HPLC.

    • Accession

      P68431
    • Animal free

      No
    • Nature

      Synthetic
      • Species

        Human
      • Modifications

        di methyl K4

    Associated products

    • Corresponding Antibody

      • Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)
    • Related Products

      • Anti-Histone H3 (di methyl K9) antibody [mAbcam 1220] - ChIP Grade (ab1220)
      • Human Histone H3 (mono methyl K4) peptide (ab1340)
      • Anti-Histone H3 (di methyl K4, tri methyl K4) antibody [mAbcam 6000] - ChIP Grade (ab6000)
      • Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
      • Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)

    Specifications

    Our Abpromise guarantee covers the use of ab7768 in the following tested applications.

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

    • Applications

      Blocking - Blocking peptide for Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

      Dot blot

    • Form

      Liquid
    • Additional notes

      - 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...

    Preparation and Storage

    • Stability and Storage

      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.

    General Info

    • Alternative names

      • 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
    • 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

    Images

    • Dot Blot - Human Histone H3 (di methyl K4) peptide (ab7768)
      Dot Blot - Human Histone H3 (di methyl K4) peptide (ab7768)This image is courtesy of an anonymous Abreview

      ab7768 (at 0.01 and 0.1 μg) used in dot blot to test for cross-reactivity of ab7766 (1/3000). An HRP-conjugated mouse anti-rabbit IgG polyclonal (1/10000) was used as the secondary antibody.

      See Abreview

    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 download

      Download

    References (6)

    Publishing research using ab7768? Please let us know so that we can cite the reference in this datasheet.

    ab7768 has been referenced in 6 publications.

    • Sun P  et al. KMT2D inhibits the growth and metastasis of bladder Cancer cells by maintaining the tumor suppressor genes. Biomed Pharmacother 115:108924 (2019). PubMed: 31100540
    • You Y  et al. Temporal dynamics of gene expression and histone marks at the Arabidopsis shoot meristem during flowering. Nat Commun 8:15120 (2017). PubMed: 28513600
    • Lin X  et al. Protocol: Chromatin immunoprecipitation (ChIP) methodology to investigate histone modifications in two model diatom species. Plant Methods 8:48 (2012). PubMed: 23217141
    • Connor C  et al. A simple method for improving the specificity of anti-methyl histone antibodies. Epigenetics 5:392-5 (2010). PubMed: 20458167
    • Vitaliano-Prunier A  et al. Ubiquitylation of the COMPASS component Swd2 links H2B ubiquitylation to H3K4 trimethylation. Nat Cell Biol 10:1365-71 (2008). PubMed: 18849979
    • Mulder KW  et al. Regulation of histone H3K4 tri-methylation and PAF complex recruitment by the Ccr4-Not complex. Nucleic Acids Res 35:2428-39 (2007). PubMed: 17392337

    Customer reviews and Q&As

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

    1-6 of 6 Abreviews or Q&A

    Other (Dot Blot) Abreview for Histone H3 peptide - di methyl K4

    Excellent
    Abreviews
    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 - di methyl K4')
    Primary antibody: Abcam primary antibody: Anti-Histone H3 (methylated) antibody - ChIP Grade (ab7766)
    Dilution: 1/3000

    Secondary antibody
    Name: Non-Abcam antibody was used: anti-rabbit
    Host species: Mouse
    Clonality: Polyclonal
    Conjugation: Horse Radish Peroxidase
    Dilution: 1/10000
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 01 2010

    Question

    I purchased a Histone H3 peptide-di methyl K4 (ab7768). This peptide is 134 amino acid long as shown on your website. However, I just couldn't detect the peptide by Western blot using anti-H3 dimethyl K4 antibody from Abcam or anti-Histone antibody (N-terminus). I am wondering if this peptide is 34 aa other than 134 aa long.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 05 2012

    Answer

    Unfortunately, I think there has been a mis-understanding. The peptide ab7768 is actually only 11 amino acids long. The histone H3 protein is 134 amino acids long, but ab7768 is a peptide derived from the protein.

    On the datasheet we specify:

    Protein description
    Synthetic peptide derived from within residues 1 - 100 of Human Histone H3, di methylated at K4. (Note: the amino acid sequence is proprietary) This peptide was used as an immunogen for ab7766 - Histone H3 (di methyl K4) antibody - ChIP Grade.

    Protein length
    134 amino acids

    Read More

    Abcam Scientific Support

    Answered on Nov 05 2012

    Question

    I am inquiring about histone H3 peptides – unmodified and modified at positions of Lys4, Lys9 and Lys27. I have checked up your products, however I cannot find the information at Abcam website about their residue sequences, residue positions and purity.
    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.
    I would like to use your products, which have been applied in several epigenomic researches with human. It will be a good reference for your company if they can be adapted successfully to Arabidopsis.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 15 2012

    Answer

    We have seven different peptides which would suit your description:




    https://www.abcam.com/Human-Histone-H3-mono-methyl-K4-peptide-ab1340.html (or use the following: https://www.abcam.com/Human-Histone-H3-mono-methyl-K4-peptide-ab1340.html).

    https://www.abcam.com/Human-Histone-H3-tri-methyl-K4-peptide-ab1342.html (or use the following: https://www.abcam.com/Human-Histone-H3-tri-methyl-K4-peptide-ab1342.html).

    https://www.abcam.com/Human-Histone-H3-mono-methyl-K9-peptide-ab1771.html (or use the following: https://www.abcam.com/Human-Histone-H3-mono-methyl-K9-peptide-ab1771.html).

    https://www.abcam.com/Human-Histone-H3-di-methyl-K9-peptide-ab1772.html (or use the following: https://www.abcam.com/Human-Histone-H3-di-methyl-K9-peptide-ab1772.html).

    https://www.abcam.com/Human-Histone-H3-tri-methyl-K9-peptide-ab1773.html (or use the following: https://www.abcam.com/Human-Histone-H3-tri-methyl-K9-peptide-ab1773.html).

    https://www.abcam.com/Human-Histone-H3-tri-methyl-K27-peptide-ab1782.html (or use the following: https://www.abcam.com/Human-Histone-H3-tri-methyl-K27-peptide-ab1782.html).

    https://www.abcam.com/Human-Histone-H3-di-methyl-K4-peptide-ab7768.html (or use the following: https://www.abcam.com/Human-Histone-H3-di-methyl-K4-peptide-ab7768.html).



    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

    Read More

    Abcam Scientific Support

    Answered on Oct 15 2012

    Question

    I’m looking to purchase peptides ab1773 and ab7768, however before I do I need to know their molecular masses to determine if we can visualise them.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 20 2012

    Answer

    The product ab1773 and ab7768 are very small peptides; due to their low molecular weight these products should not be used in SDS PAGE experiments.
    We have tested these product for blocking the antibody active sites so these products should be used in blocking experiments or in applications as recommended on the datasheet.

    Read More

    Abcam Scientific Support

    Answered on Jan 20 2012

    Question

    Can you tell me what is the molecular weight of this product?

    Read More

    Abcam community

    Verified customer

    Asked on May 14 2009

    Answer

    Thank you for your enquiry. I can confirm that the molecular weight of this product is 1390.6 Dalton.

    Read More

    Abcam Scientific Support

    Answered on May 14 2009

    Question

    I am writing to find out how the specificity of the antibodies I purchased were determined. I bought: Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895) Histone H3 (tri methyl K4) antibody - ChIP Grade (ab8580) Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766) I also purchased these peptides: Histone H3 peptide (1-8) - tri methyl K4 (ab1342) Histone H3 peptide (1-10) - di methyl K4 (ab7768) Histone H3 peptide (1-8) - mono methyl K4 (ab1340) Histone H3 peptide (1-16) - unmodified (ab7228) I would like to know of any manual dot-blotting methods you have, that will work with these. I have tried to use them in a blot to no avail so far. The proteins did not bind to the PVDF membrane I used. I was attempting a procedure from the millipore company instructions. there is no dot-blotting apparatus in my lab, so I have to do it by hand.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 06 2005

    Answer

    Thank you for your enquiry regarding Dotblots with our antibodies. Unfortunately we do not have a recommended protocol for Dot-blotting but can recommend trying a nitrocellulose membrane, a general manual will be able to help you. Specificity of those antibodies was tested by WB of Histone (ab121) (0.5µg per lane) and with blocking peptides (modified and non modified peptides). In each case, blocking was achieved only when the adequate peptide was added.

    Read More

    Abcam Scientific Support

    Answered on Apr 07 2005

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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