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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M | PIA, IF, DB, ChIP-seq, WB, ChIP | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 05-1242 |
| Description | Anti-trimethyl-Histone H3 (Lys9) Antibody, clone 6F12-H4 |
| Alternate Names |
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| 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 |
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| Presentation | Purified antibody supplied at 1 mg/mL in 0.1 M Tris-glycine, pH 7.4, 150 mm NaCl, 0.05% NaN3. |
| Quality Level | MQ100 |
| 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 |
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| 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 |
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| 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 |
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| 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 |
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| Modifications |
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| Purification Method | Purified |
| UniProt Number |
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| 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 |
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| 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 |