Key features and details
- Assay type: Cell-based (quantitative)
- Platform: Microplate reader
- Assay time: 3 hr
- Sample type: Adherent cells
Product nameHistone H3 (methyl K9) Assay Kit (In Situ)
See all Histone H3 kits
Sample typeAdherent cells
Assay typeCell-based (quantitative)
Assay time3h 00m
Species reactivityReacts with: Mouse
Predicted to work with: Mammals
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.
Histone H3 (methyl K9) Assay Kit (In Situ) (ab115047) allows the user to specifically measure histone H3K9me in situ using cultured adherent cells in a quick and efficient procedure which can be finished within 3 hours.
Storage instructionsPlease refer to protocols.
Components 96 tests 2 x 96 tests 10X Wash Buffer 1 x 30ml 2 x 30ml 30% H2O2 Solution 1 x 0.5ml 1 x 1ml 8-Well Control Strips 2 units 4 units Antibody Buffer 1 x 15ml 1 x 20ml Blocking Buffer 1 x 20ml 2 x 20ml Capture Antibody, 100 µg/mL 1 x 60µl 1 x 120µl Detection Antibody, 400 µg/mL 10 x 20µl 1 x 40µl Developing Solution 1 x 12ml 1 x 24ml Methylated H3K9 Control, 20 µg/mL 1 x 15µl 1 x 30µl Microplates 1 unit 2 units Permeabilizing Buffer 1 x 30ml 2 x 30ml Stop Solution 1 x 6ml 1 x 12ml
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 (K4) Methyltransferase Activity Quantification Assay Kit (ab113452)
- Histone H3 (K9) Methyltransferase Activity Quantification Assay Kit (ab113453)
- DNA demethylase (total) Activity Quantification Assay Kit (ab113472)
- Histone H3 (methyl K9) Assay Kit (In Situ) (ab115047)
- Histone H3 (methyl K4) Assay Kit (ab115048)
- Histone H3 (methyl K9) Assay Kit (ab115049)
- Histone H3 (methyl K27) Assay Kit (ab115050)
ab115047 has not yet been referenced specifically in any publications.