Human Histone H3 (di methyl K4) peptide (ab7768)

Reviews (1)Q&A (5)References (3)

Description

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.
  • 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

References

This product has been referenced in:

  • You Y  et al. Temporal dynamics of gene expression and histone marks at the Arabidopsis shoot meristem during flowering. Nat Commun 8:15120 (2017). Read more (PubMed: 28513600) »
  • Vitaliano-Prunier A  et al. Ubiquitylation of the COMPASS component Swd2 links H2B ubiquitylation to H3K4 trimethylation. Nat Cell Biol 10:1365-71 (2008). Read more (PubMed: 18849979) »
See all 3 Publications for this product

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

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

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.

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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

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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.

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Answer

We have seven different peptides which would suit your description:




https://www.abcam.com/index.html?datasheet=1340 (or use the following: https://www.abcam.com/index.html?datasheet=1340).

https://www.abcam.com/index.html?datasheet=1342 (or use the following: https://www.abcam.com/index.html?datasheet=1342).

https://www.abcam.com/index.html?datasheet=1771 (or use the following: https://www.abcam.com/index.html?datasheet=1771).

https://www.abcam.com/index.html?datasheet=1772 (or use the following: https://www.abcam.com/index.html?datasheet=1772).

https://www.abcam.com/index.html?datasheet=1773 (or use the following: https://www.abcam.com/index.html?datasheet=1773).

https://www.abcam.com/index.html?datasheet=1782 (or use the following: https://www.abcam.com/index.html?datasheet=1782).

https://www.abcam.com/index.html?datasheet=7768 (or use the following: https://www.abcam.com/index.html?datasheet=7768).



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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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.

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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.

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

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Abreviews
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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

Abcam user community

Verified customer

Submitted Nov 01 2010

Question

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

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Answer

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

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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.

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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.

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