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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | FC, ICC, IF, PIA, WB, Mplex | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 05-806 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-phospho-Histone H3 (Ser10) Antibody, clone 3H10 |
| Alternate Names |
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| Background Information | Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 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. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Purified mouse IgG1k in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl and 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-phospho-Histone H3 (Ser10) Antibody, clone 3H10 is a mouse monoclonal antibody for detection of Histone H3 phosphorylated at serine 10. This purified mAb, also known as H3S10p, is published in peer reviewed journals and has been validated in FC, ICC, IF, PIA, WB, Mplex. |
| Key Applications |
|
| Application Notes | Flow Cytometry: This antibody has been reported by an independent laboratory to detect phosphorylated histone H3 using flow cytometry. Beadlyte® Histone-Peptide Specificity Assay: 1:1,000 to 1:81,000 dilutions of a previous lot were incubated with histone H3 peptides containing various modifications conjugated to Luminex® microspheres. (Figure B). Only the peptide containing phospho-serine 10 was detected. Immunocytochemistry: 0.2 μg/mL of a previous lot showed positive chromosome immunostaining for mitotic A431 and HeLa cells fixed with 95% ethanol and 5% acetic acid and permeabilized with 0.1% Triton® X-100. Peptide Inhibition Analysis: Detection of histone H3 in immunoblots of colcemid treated HeLa acid extracts by anti-phospho-Histone H3 (Ser10) was diminished by 10 μM of histone H3 peptide containing phospho-serine 10, but not by peptides containing phospho-serine 28 or an unmodified histone H3 sequence (Figure C). Immunofluorescence: |
| Biological Information | |
|---|---|
| Immunogen | A proprietary immunogen based on a peptide sequence containing phospho-serine corresponding to residue 10 of human histone H3. |
| Clone | 3H10 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | Histone H3 phosphorylated at Ser10. |
| Isotype | IgG1κ |
| Species Reactivity |
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| Species Reactivity Note | Human. Broad species cross-reactivity is expected. |
| 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. |
| Gene Symbol |
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| Modifications |
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| Purification Method | Protein g Purified |
| 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 acid extracted proteins from mitotic HeLa cells treated with colcemid (Catalog # 17-306). Western Blot Analysis: 0.05-0.5 μg/mL of this lot detected phosphorylated histone H3 in acid extracted proteins from mitotic HeLa cells treated with colcemid (Catalog # 17-306), but not unmodified recombinant Histone H3 (Catalog # 14-494) (Figure A). |
| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Stable for 1 year at 2 to 8°C from date of receipt. Handling Recommendations: Upon receipt, 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: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |