Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome, the basic subunit of chromatin. Histone modifications regulate DNA transcription, repair, recombination, and replication. These modifications can alter local chromatin architecture, or recruit trans-acting factors that recognize specific histone modifications (the "histone code" hypothesis). Trimethylation of histone H3 on Lys9 (H3K9me3) is one of the most highly studied epigenetic marks. H3K9me3 functions in the repression of euchromatic genes, and in epigenetic control of heterochromatin assembly, most likely via acting as a recognition motif for the binding of chromatin-associated proteins, such as Swi6 or HP1α/β. Phosphorylated Ser10 is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition, Ser10 phosphorylation is also an essential regulatory mechanism for neoplastic cell transformation.
Use Anti-trimethyl(Lys9)-phospho(Ser10)-Histone H3 Antibody (Rabbit Monoclonal Antibody) validated in WB, PIA, DB, Mplex to detect trimethyl(Lys9)-phospho(Ser10)-Histone H3 also known as H3K9me3S10P.
Key Applications
Western Blotting
Multiplexing
Dot Blot
Peptide Inhibition Assay
Application Notes
Peptide Dot Blot Analysis: 1:500 to 1:1,000 dilutions of this antibody only detected histone H3 peptides containing the dual modifications of trimethyl-lysine/phosphoserine at residues 9/10. Peptides containing single or other dual modifications were not detected.
Beadlyte® Histone Peptide Assay: 1:1,000 to 1:2,000 dilutions of this antibody only detected histone H3 peptides containing the dual modifications of trimethyl-lysine/phosphoserine at residues 9/10.
Peptide Inhibition Analysis: 1 μM of histone H3 peptide containing trimethyl-lysine 9/phospho-serine 10 abolished detection of histone H3 by anti-trimethyl (Lys9)-phospho (Ser10)-Histone H3 (1:20,000 dilution) in immunoblots of colcemid-treated HeLa acid extracts.
Biological Information
Immunogen
KLH-conjugated, synthetic peptide containing the sequence …ARme3KpST… in which me3K corresponds to trimethyl-lysine 9 and pS to phosphoserine 10 of human histone H3.
Epitope
trimethyl (Lys9)-phospho (Ser10)
Host
Rabbit
Specificity
Recognizes histone H3 containing the dual modifications of trimethylation of Lys9 and phosphorylation of Ser10. Does not detect single modifications or other dual modifications of histone H3.
Isotype
IgG
Species Reactivity
Human
Species Reactivity Note
Human. Broad species cross-reactivity is expected, based on sequence homology.
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq]
FUNCTION:SwissProt: Q16695 # 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.
SIZE: 136 amino acids; 15508 Da SUBUNIT:The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. SUBCELLULAR LOCATION:Nucleus. TISSUE SPECIFICITY:Expressed in testicular cells. DEVELOPMENTAL STAGE:Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. PTM:Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity). SIMILARITY:SwissProt: Q16695 ## Belongs to the histone H3 family
Product Usage Statements
Quality Assurance
Evaluated by Western Blot on HeLa cells.
Western Blotting Analysis: A 1:1000-1:2000 dilution of this antibody detected trimethyl-lysine9-phosphoserine 10 Histone H3 in acid-extracted proteins from HeLa cells treated with colcemid (Figure A).
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20ºC from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Histones are highly conserved proteins that serve as the structural scaffold for the organization of nuclear DNA into chromatin. The four core histones, H2A, H2B, H3, and H4, assemble into an octamer (2 molecules of each). Subsequently, 146 base pairs of DNA are wrapped around the octamer, forming a nucleosome, the basic subunit of chromatin. Histone modifications regulate DNA transcription, repair, recombination, and replication. These modifications can alter local chromatin architecture, or recruit trans-acting factors that recognize specific histone modifications (the "histone code" hypothesis). Trimethylation of histone H3 on Lys9 (H3K9me3) is one of the most highly studied epigenetic marks. H3K9me3 functions in the repression of euchromatic genes, and in epigenetic control of heterochromatin assembly, most likely via acting as a recognition motif for the binding of chromatin-associated proteins, such as Swi6 or HP1α/β. Phosphorylated Ser10 is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition, Ser10 phosphorylation is also an essential regulatory mechanism for neoplastic cell transformation.
Use Anti-trimethyl(Lys9)-phospho(Ser10)-Histone H3 Antibody (Rabbit Monoclonal Antibody) validated in WB, PIA, DB, Mplex to detect trimethyl(Lys9)-phospho(Ser10)-Histone H3 also known as H3K9me3S10P.
Key Applications
Western Blotting
Multiplexing
Dot Blot
Peptide Inhibition Assay
Application Notes
Peptide Dot Blot Analysis: 1:500 to 1:1,000 dilutions of this antibody only detected histone H3 peptides containing the dual modifications of trimethyl-lysine/phosphoserine at residues 9/10. Peptides containing single or other dual modifications were not detected.
Beadlyte® Histone Peptide Assay: 1:1,000 to 1:2,000 dilutions of this antibody only detected histone H3 peptides containing the dual modifications of trimethyl-lysine/phosphoserine at residues 9/10.
Peptide Inhibition Analysis: 1 μM of histone H3 peptide containing trimethyl-lysine 9/phospho-serine 10 abolished detection of histone H3 by anti-trimethyl (Lys9)-phospho (Ser10)-Histone H3 (1:20,000 dilution) in immunoblots of colcemid-treated HeLa acid extracts.
Biological Information
Immunogen
KLH-conjugated, synthetic peptide containing the sequence …ARme3KpST… in which me3K corresponds to trimethyl-lysine 9 and pS to phosphoserine 10 of human histone H3.
Epitope
trimethyl (Lys9)-phospho (Ser10)
Host
Rabbit
Specificity
Recognizes histone H3 containing the dual modifications of trimethylation of Lys9 and phosphorylation of Ser10. Does not detect single modifications or other dual modifications of histone H3.
Isotype
IgG
Species Reactivity
Human
Species Reactivity Note
Human. Broad species cross-reactivity is expected, based on sequence homology.
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq]
FUNCTION:SwissProt: Q16695 # 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.
SIZE: 136 amino acids; 15508 Da SUBUNIT:The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. SUBCELLULAR LOCATION:Nucleus. TISSUE SPECIFICITY:Expressed in testicular cells. DEVELOPMENTAL STAGE:Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. PTM:Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity). SIMILARITY:SwissProt: Q16695 ## Belongs to the histone H3 family
Product Usage Statements
Quality Assurance
Evaluated by Western Blot on HeLa cells.
Western Blotting Analysis: A 1:1000-1:2000 dilution of this antibody detected trimethyl-lysine9-phosphoserine 10 Histone H3 in acid-extracted proteins from HeLa cells treated with colcemid (Figure A).
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20ºC from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
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