Overview

  • Product name

    Histone H3 (methyl K9) Assay Kit
    See all Histone H3 kits
  • Sample type

    Tissue, Adherent cells, Suspension cells
  • Assay type

    Quantitative
  • Assay time

    5h 00m
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Mammals
  • Product overview

    Histone H3 (methyl K9) Assay Kit (ab115049) allows the user to specifically measure global histone H3K9 methylation using a variety of mammalian cells including fresh and frozen tissues, cultured adherent and suspension cells.


    Methylation of histone H3 at lysine 9 (K9) mediates heterochromatin formation by forming a binding site for HP1 and also participates in silencing expression at euchromatic sites. Moreover, presence of H3 (methyl K9) positively correlates with DNA methylation.


     

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 48 tests 96 tests
    10X Lysis Buffer 1 x 5ml 1 x 10ml
    10X Wash Buffer 1 x 14ml 1 x 28ml
    8-Well Assay Strips (with Frame) 6 units 12 units
    Antibody Buffer 1 x 6ml 1 x 12ml
    Blocking Buffer 1 x 10ml 1 x 20ml
    Capture Antibody, 100 µg/mL 1 x 25µl 1 x 50µl
    Detection Antibody, 400 µg/mL 1 x 10µl 1 x 20µl
    Developing Solution 1 x 5ml 1 x 10ml
    Extraction Buffer 1 x 8ml 1 x 16ml
    Histone Buffer 1 x 0.5ml 1 x 1ml
    Methylated H3K9 Control, 60 µg/mL 1 x 10µl 1 x 20µl
    Stop Solution 1 x 3ml 1 x 6ml
  • Research areas

  • 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
  • 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
  • Database links

Protocols

References

ab115049 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Answer

Thank you for your enquiry.

I can confirm that the ab115049 EpiSeeker Histone H3 (methyl K9) Assay Kit is shipped on ice at 4oC. However, please note that you will need to check the storage conditions for each individual kit component once it is delivered and store them as directed as soon as possible. This information can be found in the kit protocol, provided under the protocol tab on the online datasheet.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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