Human Histone H3 (acetyl K9) peptide (ab16635)

Reviews (1)Q&A (3)References (2)

Key features and details

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

Description

Associated products

Specifications

Our Abpromise guarantee covers the use of ab16635 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 (acetyl K9) antibody - ChIP Grade (ab10812)

  • Form

    Lyophilized

  • 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.

  • Reconstitution
    Reconstitute with 100 ul of deionised water (or equivalent).

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

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

References (2)

Publishing research using ab16635? Please let us know so that we can cite the reference in this datasheet.

ab16635 has been referenced in 2 publications.

  • Vasudevarao MD  et al. Naphthoquinone-mediated inhibition of lysine acetyltransferase KAT3B/p300, basis for non-toxic inhibitor synthesis. J Biol Chem 289:7702-17 (2014). PubMed: 24469461
  • Dalvoy Vasudevarao M  et al. Inhibition of acetyltransferase alters different histone modifications: probed by small molecule inhibitor plumbagin. J Biochem 152:453-62 (2012). PubMed: 22923743

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Question

What is the exact molecular weight of this peptide? The sequence is:
ARTKQTARK(Ac)STGG-C
Mass spec results are not confirming the expected molecular weight of this sequence. Is there any group on the N-terminus? Is there a C-terminal protection group on the cysteine, or just -COOH?

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Answer

I've attached the mass spec results for this peptide, which were obtained by the lab that produced the peptide.

The average MW of the peptide is 1505.7, and the ESI-TOF-MS is 1504.9. There is a C-terminal amide and the acetyled lysine, but no other modifications.

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Question

We would like to know the Molecular Weight of H3K9, which we have already bought.

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Answer

Based on the peptide sequence, this peptide should be ˜12kDA before modification. As this has been acetylated is may be 1 or 2 kDa larger.

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Other (Dot Blot) Abreview for Histone H3 peptide - acetyl K9

 Excellent
Abreviews
Application
Other

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 - acetyl K9')
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

Abcam user community

Verified customer

Submitted Nov 04 2010

Question

Can this product ab16635 be used as a blocking peptide for ab12178 in ChIP?

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Answer

Thank you for your enquiry. Blocking peptides could be used prior to the ChIP procedure to inhibit the binding capacity of the antibody. ab16635 Histone H3 peptide - acetyl K9 would be the appropriate peptide for ab12178 Histone H3 (acetyl K9) antibody. I would suggest incubating the antibody with the peptide prior to adding to the chromatin/protein preparation for the immunoprecipitation. I would suggest a starting concentration of approximately 1ug/ml.

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