Anti-trimethyl Histone H3(Lys9), clone 6F12-H4 (mouse monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
H, M	PIA, IF, DB, ChIP-seq, WB, ChIP	M	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

H, M

PIA, IF, DB, ChIP-seq, WB, ChIP

Purified

Monoclonal Antibody

Description
Catalogue Number	05-1242
Description	Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4
Alternate Names	H3K9me3 Histone H3 (tri methyl K9) H3 histone family, member T histone 3 H3 histone cluster 3 H3
Background Information	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. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. 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. The enzymes responsible for H3K9me3 formation are SUV39H1 and SUV39H2.

Description

Catalogue Number

05-1242

Description

Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4

Alternate Names

H3K9me3
Histone H3 (tri methyl K9)
H3 histone family, member T
histone 3
H3
histone cluster 3
H3

Background Information

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. The most commonly studied modifications are acetylation, phosphorylation, methylation, and ubiquitination. 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. The enzymes responsible for H3K9me3 formation are SUV39H1 and SUV39H2.

Product Information
Format	Purified
Control	HeLa Acid Extract
Presentation	Purified antibody supplied at 1 mg/ml in 0.1 M Tris-glycine, pH 7.4, 150 mM NaCl, 0.05% NaN3.

Product Information

Format

Purified

Control

HeLa Acid Extract

Presentation

Purified antibody supplied at 1 mg/ml in 0.1 M Tris-glycine, pH 7.4, 150 mM NaCl, 0.05% NaN3.

Applications
Application	Use Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4 (mouse monoclonal antibody) validated in ChIP, PIA, IF, DB, ChIP-seq, WB to detect trimethyl-Histone H3 (Lys9) also known as H3K9me3, Histone H3 (tri methyl K9).
Key Applications	Peptide Inhibition Assay Immunofluorescence Dot Blot ChIP-seq Western Blotting Chromatin Immunoprecipitation (ChIP)
Application Notes	Chromatin Immunoprecipitation (ChIP): Representative data from a previous lot. Sonicated 3T3 L1 chromatin was subjected to chromatin immunoprecipitation using anti- trimethyl-histone H3 (Lys9) and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of trimethylhistone H3 (Lys9) associated DNA fragments was verified by qPCR using primers flanking the p16 promoter. Peptide Inhibition Analysis: Peptide blocking assay demonstrates distinct preference of the antibody for the trimethyl form vs. the dimethyl form. Chromatin Immunoprecipitation (ChIP): ChIP analysis of known chromosomal Suv39h targets (H3K9me3 in major satellites, mouseES cells). Dot Blot Analysis: Dot-blot analysis demonstrating specificity of anti-H3K9me3, clone 6F12-H4 for trimethyl Lys9 of histone H3.

Applications

Application

Use Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4 (mouse monoclonal antibody) validated in ChIP, PIA, IF, DB, ChIP-seq, WB to detect trimethyl-Histone H3 (Lys9) also known as H3K9me3, Histone H3 (tri methyl K9).

Key Applications

Peptide Inhibition Assay
Immunofluorescence
Dot Blot
ChIP-seq
Western Blotting
Chromatin Immunoprecipitation (ChIP)

Application Notes

Chromatin Immunoprecipitation (ChIP):
Representative data from a previous lot. Sonicated 3T3 L1 chromatin was subjected to chromatin immunoprecipitation using anti- trimethyl-histone H3 (Lys9) and the Magna ChIP G (Cat. #17-611) Kit. Successful immunoprecipitation of trimethylhistone H3 (Lys9) associated DNA fragments was verified by qPCR using primers flanking the p16 promoter.

Peptide Inhibition Analysis:
Peptide blocking assay demonstrates distinct preference of the antibody for the trimethyl form vs. the dimethyl form.

Chromatin Immunoprecipitation (ChIP):
ChIP analysis of known chromosomal Suv39h targets (H3K9me3 in major satellites, mouseES cells).

Dot Blot Analysis:
Dot-blot analysis demonstrating specificity of anti-H3K9me3, clone 6F12-H4 for trimethyl Lys9 of histone H3.

Biological Information
Immunogen	KLH-conjugated, branched synthetic peptide containing the sequence (QTARme3K- STGGKA)2-KC, in which me3K corresponds to trimethyl lysine 9 of human histone H3. Data suggests this antibody may require the presence of the glutamine residue in position 5 for binding.
Epitope	Trimethyl Lys9
Clone	6F12-H4
Host	Mouse
Specificity	Recognizes trimethyl-histone H3 (Lys9), Mr 17 kDa. Cross-reacts with dimethyl histone H3(Lys9) at higher concentrations.
Isotype	IgG1κ
Species Reactivity	Human Mouse
Species Reactivity Note	Human and mouse. Based on sequence homology, broad species cross-reactivity is expected with all mammals, Drosophila, Xenopus, and Arabidopsis.
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NM_003493
Entrez Gene Summary	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]
Gene Symbol	H3.4 H3/g H3/t H3FT H3t MGC126886 OTTHUMP00000037945
Modifications	Methylation
Purification Method	Purified
UniProt Number	Q16695
UniProt Summary	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. 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. 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: Belongs to the histone H3 family.
Molecular Weight	~17 kDa

Biological Information

Immunogen

KLH-conjugated, branched synthetic peptide containing the sequence (QTARme3K- STGGKA)2-KC, in which me3K corresponds to trimethyl lysine 9 of human histone H3. Data suggests this antibody may require the presence of the glutamine residue in position 5 for binding.

Epitope

Trimethyl Lys9

Clone

6F12-H4

Host

Mouse

Specificity

Recognizes trimethyl-histone H3 (Lys9), Mr 17 kDa. Cross-reacts with dimethyl histone H3(Lys9) at higher concentrations.

Isotype

IgG1κ

Species Reactivity

Human
Mouse

Species Reactivity Note

Human and mouse. Based on sequence homology, broad species cross-reactivity is expected with all mammals, Drosophila, Xenopus, and Arabidopsis.

Antibody Type

Monoclonal Antibody

Entrez Gene Number

NM_003493

Entrez Gene Summary

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]

Gene Symbol

H3.4
H3/g
H3/t
H3FT
H3t
MGC126886
OTTHUMP00000037945

Modifications

Methylation

Purification Method

Purified

UniProt Number

Q16695

UniProt Summary

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.

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.

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: Belongs to the histone H3 family.

Molecular Weight

~17 kDa

Product Usage Statements
Quality Assurance	Routinely evaluated by Western Blot on HeLa acid extracts. Western Blot Analysis: A 0.5 – 5 μg dilution of this lot detected trimethyl histone H3 (Lys9) in HeLa acid extracts.
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.

Product Usage Statements

Quality Assurance

Routinely evaluated by Western Blot on HeLa acid extracts.

Western Blot Analysis: A 0.5 – 5 μg dilution of this lot detected trimethyl histone H3 (Lys9) in HeLa acid extracts.

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 2-8°C from date of receipt. For maximum recovery of product, centrifuge the vial prior to removing the cap. Avoid freezing, which can damage IgG and affect product performance.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at 2-8°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap. Avoid freezing, which can damage IgG and affect product performance.

Packaging Information
Material Size	200 µL

Packaging Information

Material Size

200 µL

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Anti-trimethyl Histone H3(Lys9), clone 6F12-H4 (mouse monoclonal)

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