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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | Mplex, WB, PIA, ChIP-seq, DB | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-539 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-acetyl-Histone H3 (Lys 4) 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. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. 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 amino-terminal tails of histone proteins are subject to posttranslational modifications, including acetylation and methylation, which recruit downstream regulatory factors, influence chromatin structure, and are critical determinants of transcription. Acetylation of histone H3 occurs at several different lysine positions in the histone tail and is performed by a family 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. There is a significant correlation between acetylation of histones H3 in promoter regions and transcriptional activity. In contrast, dimethylation of histone H3 Lys 4 in coding regions correlates with transcriptional activity. (Ref.: Duan, MR., and Smerdon, MJ (2014). J. Biol. Chem. 289(12); 8353-8363; Nicolas, E., et Al. (2003). Mol. Cell. Biol. 23(5); 1614-1622; Bernstein, BE., et Al. (2002). Proc. Natl. Acad. Sci. USA. 99(13);8695-8700). |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Presentation | Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mm NaCl with 0.05% sodium azide with 30% glycerol. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-acetyl-Histone H3 (Lys4), Cat. No. 07-539, is a rabbit polyclonal antibody that detects Histone H3 acetylated on Lysine 4. |
| Key Applications |
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| Application Notes | Tested Applications Immunocytochemistry Analysis: A 1:200 dilution from a representative lot detected Acetyl-Histone H3 (Lys4) in HepG2 cells. Peptide Inhibition Assay: Target band detection in acid extract of HeLa cells treated with sodium butyrate was prevented by preblocking of a representative lot with Acetyl-Histone H3 (Lys4) peptide, but not with unmodified peptide or with Acetyl-Histone H3 (Lys9) peptide. DOT Blot Analysis: Anti-acetyl-Histone H3 (Lys4) antibody, Cat. No. 07-539 detected 4 ng of Acetyl-Histone H3 (Lys4) peptide, but did not detect un-modified Histone H3 peptide, or Histone H3 peptides acetylated on (Lys9), (Lys14), (Lys18), and (Lys23). Chromatin Immunoprecipitation (ChIP) Analysis: 1 µg of this antibody detected Acetyl-Histone H3 (Lys4) in Sonicated chromatin prepared from HeLa cells ( 1X106 IP equivalent) . 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 10 amino acids from the N-terminal region of human Histone H3 surrounding acetylated lysine 4. |
| Epitope | N-terminal |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody specifically detects Histone H3 acetylated on Lysine 4. |
| Isotype | IgG |
| Species Reactivity |
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| Species Reactivity Note | Human. Predicted to react with All species 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 nucleosomes 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. |
| Gene Symbol |
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| Modifications |
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| Purification Method | Affinity 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. 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 | ~16 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 of HeLa cells treated with Sodium Butyrate. Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected Acetyl-Histone H3 (Lys4) in acid extract of HeLa cells treated with Sodium Butyrate ( 5 mm; 24 H), but not in untreated HeLa cells. |
| 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 |