Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 2 hr 30 min
- Sample type: Adherent cells, Suspension cells, Tissue
Product nameHistone H3 (di-methyl K9) Quantification Kit (Fluorometric)
See all Histone H3 kits
Sample typeTissue, Adherent cells, Suspension cells
Assay time2h 30m
Species reactivityReacts with: Mouse, Human
Predicted to work with: Mammals
Methylation of histone H3 at lysine 9 by ESET, G9a among other methyl transferases is involved in euchromatin gene silencing as well as in heterochromatin formation. Specifically, H3 (di-methyl K9) mediates heterochromatin formation by forming a binding site for HP1.
Histone H3 (di-methyl K9) Quantification Kit (Fluorometric) (ab115063) allows the user to specifically measure global di-methylation of histone H3K9 fluorometrically using a variety of mammalian cells (human, mouse, etc.) including fresh and frozen tissues, cultured adherent and suspension cells.
Storage instructionsPlease refer to protocols.
Components Identifier 48 tests 96 tests 10X Wash Buffer 1 x 10ml 1 x 20ml 8-Well Sample Strips (with Frame) 4 units 9 units 8-Well Standard Control Strips Green Ringed 2 units 3 units Antibody Buffer 1 x 6ml 1 x 12ml Detection Antibody, 1 mg/mL 1 x 5µl 1 x 10µl Fluoro Developer 1 x 12µl 1 x 24µl Fluoro Dilution 1 x 4ml 1 x 8ml Fluoro Enhancer 1 x 12µl 1 x 24µl Standard Control, 100 µg/mL 1 x 10µl 1 x 20µl
FunctionCore 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 similaritiesBelongs to the histone H3 family.
Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
modificationsAcetylation 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 localizationNucleus. Chromosome.
- Information by UniProt
- H3 histone family member E pseudogene
- H3 histone family, member A
- Histone H3 (K9) Methyltransferase Activity Quantification Assay Kit (ab113453)
- Histone H3 (methyl K9) Assay Kit (In Situ) (ab115047)
- Histone H3 (mono-methyl K9) Quantification Kit (Colorimetric) (ab115060)
- Histone H3 (mono-methyl K9) Quantification Kit (Fluorometric) (ab115061)
- Histone H3 (di-methyl K9) Quantification Kit (Colorimetric) (ab115062)
- Histone H3 (tri-methyl K9) Quantification Kit (Colorimetric) (ab115064)
- Histone H3 (tri-methyl K9) Quantification Kit (Fluorometric) (ab115065)
- Histone H3 (pan-methyl K9) Quantification Kit (Colorimetric) (ab115066)
- Histone H3 (pan-methyl K9) Quantification Kit (Fluorometric) (ab115067)
Histone extracts were prepared from MDA-MB-231 cells using the Histone Extraction Kit (ab113476) and the amount of H3K9me2 was measured using ab115063.
ab115063 has not yet been referenced specifically in any publications.