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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| M, H, Eu | ICC, WB, ChIP-seq, ChIP | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-081 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-phospho (Ser10)-acetyl (Lys14)-Histone H3 Antibody |
| Alternate Names |
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| Background Information | Histone H3.1 (UniProt: Q16695; also known as H3K14AcS10P, Histone H3 (acetyl K14, phospho S10), H3 histone family member T, Histone 3, H3, Histone cluster 3, H3) is encoded by the HIST3H3 (also known as HIST1H3A, H3FA, HIST1H3B, H3FL, HIST1H3C, H3FC, HIST1H3D, H3FB, HIST1H3E, H3FD, HIST1H3F, H3FI, HIST1H3G, H3FH, HIST1H3H, H3FK, HIST1H3I, H3FF, HIST1H3J, H3FJ, HIST3H3, H3FT, H3T, MGC126886) gene (Gene ID: 8350, 8351, 8352, 8353, 8354, 8355, 8356, 8357, 8358, 8968) in human. Histone H3 has two main variants, H3.1 and H3.3, which show different genomic localization patterns in animals. The H3.1 and H3.3 complexes also possess distinct histone chaperones, CAF-1 and HIRA, which play important role in mediating DNA-synthesis-dependent and -independent nucleosome assembly. It has been reported that Histone H3.1 serves as the canonical histone, which is incorporated during DNA replication, whereas H3.3 acts as the replacement histone that can be incorporated outside of S-phase during chromatin-disrupting processes like transcription. Histone H 3.1 is a core component of nucleosome that is present only in mammals and is usually enriched in acetylation of Lysine 15 and demethylation of lysine 10 (HeK9Me2). It is expressed during S phase, then expression decreases significantly as cell division slows down during the process of differentiation. Histone H 3.1 expression is shown to be replication dependent. It s presence at the site of UV-induced DNA damage has also been reported. It has also been shown that H3.1/H4 tetramers do not split and remain intact during replication dependent deposition of H3.1 variant. Phosphorylation at serine 10 by aurora kinase B is reported to mediate the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin and this phosphorylation is considered to be an essential regulatory mechanism for neoplastic cell transformation. Acetylation of lysine 14 of histone H3 (H3K14ac) is shown to be critical for DNA damage checkpoint activation by directly regulating the compaction of chromatin and by recruiting chromatin remodeling protein complex RSC (Remodels the Structure of Chromatin). Nucleosomes with H3K14ac display higher affinity for purified RSC. (Ref.: Stroud, H., et Al (2012). Proc. Natl. Acad. Sci. USA 109(14); 5370-75; Dunn, MR., and Smerdon MJ (2014). J. Biol. Chem. 289(12);8353-63).. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
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| Presentation | Purified rabbit polyclonal IgG in buffer containing 0.2 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide, before the addition of glycerol to 30%. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-phospho (Ser10)-acetyl (Lys14)-Histone H3, Cat. No. 07-081, is a highly specific rabbit polyclonal antibody that targets Histone H3 phosphorylated on Serine 10 and acetylated on Lysine 14 and has been tested for use in Immunocytochemistry, Peptide Inhibition Assay, ChIP-seq and Chromatin Immuno |
| Key Applications |
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| Application Notes | Western Blot Analysis: 1:2000-1:10,000 dilutions of a previous lot detected phosphorylated and acetylated histone H3 in acid extracted proteins from serum starved mouse 10T1/2 cells treated with 50ng/mL EGF for 12.5 minutes. Western blotting of this lot to acid extracted proteins described above was preferentially competed by peptides in the following order: phospho-acetyl-H3 (7-20) > straight chain H3 (7-20) > phospho-H3 (7-20) or acetyl-H3 (7-20). Peptide Inhibition Analysis: A 1:500 dilution from a representative lot was used with HeLa AE for peptide block analysis. Chromatin Immunoprecipitation: A previous lot of this antibody was reported by an independent laboratory to immunoprecipitate chromatin. (Cheung, P., 2000.) Immunocytochemistry: A previous lot of this antibody detected phosphorylated and acetylated histone H3 in EGF-stimulated cells; as reported by an independent laboratory. (Cheung, P., 2000.) |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated, synthetic peptide corresponding to amino acids 7-20 of human histone H3 (ARK[PS]TGGAcKAPRKQL-C). |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | Recognizes Histone H3 phosphorylated at serine 10 and acetylated at lysine 14, Mr 17 kDa. An unidentified protein is also seen in some cells lines, Mr 40 kDa. |
| Isotype | IgG |
| Species Reactivity |
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| Species Reactivity Note | Human and mouse; broad species cross-reactivity expected. |
| 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. 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. |
| Gene Symbol |
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| Modifications |
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| Purification Method | Immunoaffinity and reverse-affinity chromatography. |
| UniProt Number |
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| UniProt Summary | 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. |
| Molecular Weight | 17 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Routinely evaluated by Western Blot on HeLa acid extract. Western Blot Analysis: 1:500 dilution of this lot detected phospho acetyl Histone H3 on 10 μg of HeLa acid extract. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, 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. Note: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |