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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| M, H | WB, ICC, PIA | Rb | Unpurified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-424 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-phospho-Histone H3 (Thr3) Antibody |
| Alternate Names |
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| Background Information | Histone H3.1t (UniProt: Q16695; also known as H3/t, H3t, H3/g) is encoded by the HIST3H3 (also known as H3FT) gene (Gene ID: 8290) in human. Histones are highly conserved basic nuclear proteins that are responsible for the nucleosome structure of chromatin in eukaryotes. They play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. 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 that require DNA as a template. Histone H3 features a main globular domain and a long N-terminal tail, which protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. The N-terminal tails of histones are subject to posttranslational modifications, including phosphorylation, acetylation, and methylation, which recruit downstream regulatory factors, influence chromatin structure, and are critical determinants of transcription. Histone H3 can undergo acetylation on several lysine residues in the histone tail by histone acetyltransferases. Acetylation is generally associated with transcriptional activity and methylation of lysine and arginine residues can either activate or repress depending on the residue modified. Trimethylation of histone H3 is one of the most studied epigenetic marks. H3K4me3 modifications are reported to occur consistently at transcription start sites and H3K4me3 domain is associated with higher transcription activity and cell identity in pre-implantation development and in the process of deriving embryonic stem cells from the inner cell mass and trophoblast stem cells from the trophectoderm. Histone H3 is phosphorylated at threonine 4 (H3T3ph) by histone H3 associated protein kinase (HASPIN) during prophase and is dephosphorylated during anaphase and phosphorylation at serine 11 (H3S10ph) by Aurora kinase B is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. (Ref.: Sharifi-Zarchi, A., et Al. (2017). BMS Genomics. 18; Article 964; Liu, X., et Al. (2016). Nature. 537(7621); 558-562). |
| Product Information | |
|---|---|
| Format | Unpurified |
| Control |
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| Presentation | Immunodepleted rabbit serum with 0.05% sodium azide with 30% glycerol. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-phospho-Histone H3 (Thr3), Cat. No. 07-424, is a rabbit polyclonal antibody that detects Histone H3 phosphorylated on Threonine 3 and is tested for use in Immunocytochemistry, Peptide Inhibition Assay, and Western Blotting. |
| Key Applications |
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| Application Notes | Tested Applications Peptide Inhibition Assay: Target band detection in acid extract from colcemid treated (50 ng/mL; overnight) HeLa cells was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not with non-phosphorylated Histone H3 peptide. Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected Phospho-Histone H3 (Thr3) in NIH3T3 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 13 amino acids surrounding phosphothreonine 3 of human Histone H3. |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody detects Histone H3 phosphorylated on threonine 3. |
| Species Reactivity |
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| Species Reactivity Note | Human, Mouse. |
| 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 | Immunodepleted |
| 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 observed; 15.51 kDa calculated. Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in acid extract from colcemid-treated HeLa cells. Western Blotting Analysis: A 1:5,000 dilution of this antibody detected Phospho-Histone H3 (Thr3) in acid extract of colcemid treated (50 ng/mL, overnight) HeLa cells, but not the recombinant Histone H3 protein. |
| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Store at -10°C to -25°C. 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. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |