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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | WB, ChIP | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 09-838 |
| Description | Anti-Histone H3.3 Antibody |
| Alternate Names |
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| Background Information | Histone H3.3 (UniProt: P84243; also known as H3.3) is encoded by the H3-3A (also known as H3.3A, H3F3, H3F3A, PP781, H3-3B) gene (Gene ID: 3020) in human. Histones are basic nuclear proteins that are responsible for the nucleosome structure of chromatin in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which DNA is wrapped in repeating units, called nucleosomes, which limits DNA accessibility to the cellular machineries, which require DNA as a template. Histone H3 has two main variants, H3.1 and H3.3, which show different genomic localization patterns in animals. Histone H3.1 serves as the canonical histone, which is incorporated during DNA replication. Histone H3.3 is a highly conserved variant form of Histone H3, which replaces conventional H3 in a wide range of nucleosomes in active genes. It constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. It is associated with actively expressed genes in both animals and plants. It is predominantly enriched near transcription end sites (TES) of genes and positively associated with transcription. Histone H3 contains a main globular domain and a long N-terminal tail and is involved with the structure of the nucleosomes of the beads on a string structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. Mutations in Histone H3.3 have been implicated in a high proportion of malignant pediatric brain cancers. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
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| Presentation | Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Histone H3.3, Cat. No. 09-838, is a rabbit polyclonal antibody that detects Histone H3.3 and is tested for use in Western Blotting and Chromatin Immunoprecipitation (ChIP). |
| Key Applications |
|
| Application Notes | Tested applications Chromatin Immunoprecipitation Analysis: 2 µg from a representative lot detected Histone H3.3 in sonicate chromatin prepared from one million equivalent of HeLa cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user. |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to 12 amino acids from the C-terminal half of human Histone H3.3. |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody detects histone H3.3. It targets an epitope within 12 amino acids from the C-terminal half. |
| Species Reactivity |
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| Species Reactivity Note | Human. Predicted to react with Bovine, Mouse based on 100% sequence homology. |
| Antibody Type | Polyclonal 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. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between cleosomes and functions in the compaction of chromatin into higher order structures. This gene contains introns and its mRNA is polyadenylated, unlike most histone genes. The protein encoded is a replication-independent member of the histone H3 family. [provided by RefSeq]. |
| Gene Symbol |
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| Purification Method | Affinity Purfied |
| UniProt Number |
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| UniProt Summary | FUNCTION: Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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. Interacts with HIRA, a chaperone required for its incorporation into nucleosomes. SUBCELLULAR LOCATION: Nucleus. DEVELOPMENTAL STAGE: Expressed throughout the cell cycle independently of DNA synthesis. PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8sme2). 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 (H3R8sme2) 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. Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5 (H3K4me), Lys-37 and Lys-80. 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), which are linked to gene repression, are underrepresented. 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. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, probably DAPK3. 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 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation on Ser-32 is specific to regions bordering centromeres in metaphase chromosomes. Ubiquitinated (By similarity). SEQUENCE SIMILARITIES: Belongs to the histone H3 family. SEQUENCE CAUTION: The sequence CAH73371.1 differs from that shown. Reason: Erroneous gene model prediction. |
| Molecular Weight | ~17 kDa observed; 15.33 kDa calculated. Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in HeLa cells acid extract. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Histone H3.3 in HeLa cells acid extract. |
| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Recommended storage: +2°C to +8°C. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |