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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, DB | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABE263 |
| Description | Anti-unmethylated-Histone H3 (Lys9) Antibody, clone 9B1-2G6 |
| Alternate Names |
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| Background Information | Histones are nuclear proteins that form octameric structures which bind DNA to form units of chromatin called nucleosomes. The family of histones—H2A, H2B, H3, and H4—are key players in gene regulation. They undergo a number of post-translational modifications (PTM) in response to various stimuli, including phosphorylation on serine and threonine residues and methylation on lysine residues. PTMs produce configural changes in histone proteins that may induce nucleosome remodeling and expose or hide DNA sequences from transcriptional complexes. Methylation occurs on histone H3 at lysine 4, 9, 27, 36, and 79 residues. Previous studies have reported that methylation of H3 lysine 9 provides a binding site for heterochromatin adaptor proteins (HP1) which are involved in the formation of heterochromatin and gene repression. Conversely,deacetylation of histone H3 lysine 9 is associated with gene-activating events. Often, it is the pattern of different modifications on several histone residues that alter chromatin, rather than modification of single residues. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Purified mouse monoclonal IgG1κ supernatant in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Use Anti-unmethylated-Histone H3 (Lys9) Antibody, clone 9B1-2G6 (Mouse Monoclonal Antibody) demonstrated to detect unmethylated-Histone H3 (Lys9) also known as Histone H3.1. |
| Key Applications |
|
| Application Notes | Western Blot Analysis: 0.5 µg/mL from a representative lot detected unmethylated Histone H3 (Lys9) in 10 µg of HeLa acid extract. Dot Blot Analysis: Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Cat. No MABE263-S, unmethylated-Histone H3 (Lys9), clone 9B1-2G6 at 0.5 µg/mL (1:2,000) dilution. Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and a chemiluminescence detection system. Dot Blot (Specificity) Analysis: A representative lot detected non-methylated Histone H3 (Lys9) in Dot Blot. This antibody detects non-methylated Histone H3 (Lys9) when Ser10 is phosphorylated. It also detects mono-/di-methyl-Histone H3(Lys4), and Acetyl-Histone H3 (Lys14). (Prof. E. Ogris, Department of Medical Biochemistry, Medical University of Vienna). |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to human unmethylated Histone H3 (Lys9). |
| Epitope | unmethylated Lys9 |
| Clone | 9B1-2G6 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | This antibody recognizes Histone H3 unmethylated at Lys9. This antibody does not cross react with methyl-Histone (Lys29). This antibody detects unmethylated Histone H3 (Lys9) when Ser10 is phosphorylated. It also detects mono-/di-methyl-Histone H3 (Lys4), and Acetyl-Histone H3 (Lys14). (Prof. E. Ogris, Department of Medical Biochemistry, Medical University of Vienna). |
| Isotype | IgG1κ |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein G 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 STRUCTURE: 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. Chromosome. DEVELOPMENTAL STAGE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. POST-TRANSLATIONAL MODIFICATION: Acetylation 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 PRKCB 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 Thr-12 (H3T11ph) by chromatin-associated CHEK1 regulates the transcription of cell cycle regulatory genes by modulating acetylation of Lys-10 (H3K9ac). Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin. 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. Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination. MISCELLANEOUS: This histone is only present in mammals. SEQUENCE SIMILARITIES: Belongs to the histone H3 family. |
| Molecular Weight | ~15 kDa observed |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blot in recombinant Histone H3. Western Blot Analysis: 0.5 µg/mL of this antibody detected unmethylated Histone H3 (Lys9) in 10 µg of recombinant Histone H3. |
| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
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| Material Size | 100 µg |